Synthesis of Triethylenetetramines

ABSTRACT

Methods and intermediates for synthesizing triethylenetetramine and salts thereof, as well as novel triethylenetetramine salts and their crystal structure, and triethylenetetramine salts of high purity.

RELATED APPLICATION

This application claims benefit of priority from U.S. provisionalapplication No. 60/589,080 filed Jul. 19, 2004, to Jona, M. et al.entitled “Synthesis of Triethylenetetramines,” the content-of whichis-hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The field relates to chemicals and methods of chemical synthesisincluding, for example, novel methods for synthesis oftriethylenetetramines and triethylenetetramine salts, and crystals andpolymorphs thereof, as well as intermediates for, or within, saidsynthesis methods. The compounds have utility in a variety oftherapeutic areas.

BACKGROUND OF THE INVENTION

The following includes information that may be useful in understandingthe present inventions. It is not an admission that any of theinformation provided herein is prior art, or relevant, to the presentlydescribed or claimed inventions, or that any publication or documentthat is specifically or implicitly referenced is prior art.

Polyethylenepolyamines include triethylenetetramines that act as copperantagonists. Triethylenetetramine, sometimes also referred to astrientine, N,N′-Bis(2-aminoethyl)-1,2-ethanediamine,1,8-diamino-3,6-diazaoctane, 3,6-diazaoctane-1,8-diamine,1,4,7,10-tetraazadecane, trien, TETA, TECZA,N,N′-Bis(aminoethyl)ethylenediamine,N,N′-Bis(2-aminoethyl)ethanediamine, andN,N′-Bis(2-aminoethyl)-ethylenediamine, is a copper chelating agent.Triethylenetetramine is used as an epoxy curing agent. Merck Index, p.9478 (10th Edition, 1983). It has also been used as a thermosettingresin, as a lubricating oil additive, and as an analytical reagent forcopper and nickel id. Triethylenetetramine dihydrochloride has also beenused for treating individuals with Wilson's disease. See, for example,id.; Dubois, R. S., Lancet 2(7676):775 (1970); Walshe, J. M., Q. J. Med.42(167):441-52 (1973); Haslam, R. H., et al., Dev. Pharmacol. Ther.1(5):318-24 (1980). It has also reportedly been used to treatindividuals with primary biliary cirrhosis. See, for example, Epstein,O., et al., Gastroenterology 78(6):1442-45 (1980). In addition,trientine has been tested for inhibition of the spontaneous developmentof hepatitis and hepatic tumors in rats. See, for example, Sone, H., etal., Hepatology 23:764-70 (1996). U.S. Pat. Nos. 6,897,243, 6,610,693and 6,348,465 describe the use of copper binding compounds in thetreatment of various disorders, including treatment of diabetes mellitusand complications thereof, including, for example, diabeticcardiomyopathy.

Trientine was said to be used in the synthesis ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminein French Patent No. FR2810035 to Guilard et al. Cetinkaya, E., et al.,“Synthesis and characterization of unusual tetraaminoalkenes,” J. Chem.Soc. 5:561-7 (1992), is said to be directed to synthesis ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminefrom trientine, as is Araki T., et al., “Site-selective derivatizationof oligoethyleneimines using five-membered-ring protection method,”Macromol., 21:1995-2001 (1988). Triethylenetetramine may reportedly alsobe used in the synthesis of N-methylated triethylenetetramine, asreported in U.S. Pat. No. 2,390,766, to Zellhoefer et al.

Synthesis of polyethylenepolyamines, including triethylenetetramines,from ethylenediamine and monoethanolamine using pelleted group IVb metaloxide-phosphate type catalysts was reported by Vanderpool et al. in U.S.Pat. No. 4,806,517. Synthesis of triethylenetetramine fromethylenediamine and ethanolamine was also proposed in U.S. Pat. No.4,550,209, to Unvert et al. U.S. Pat. No. 5,225,599, to King et al. issaid to be directed to the synthesis of linear triethylene tetramine bycondensation of ethylenediamine and ethylene glycol in the presence of acatalyst. Joint production of triethylenetetramine and1-(2-aminoethyl)-aminoethyl-piperazine was proposed by Borisenko et al.in U.S.S.R. Patent No. SU1541204. U.S. Pat. No. 4,766,247 and EuropeanPatent No. EP262562, both to Ford et al., reported the preparation oftriethylenetetramine by reaction of an alkanolamine compound, analkaline amine and optionally either a primary or secondary amine in thepresence of a phosphorous containing catalyst, for example phosphoricacid on silica-alumina or Group IIIB metal acid phosphate, at atemperature from about 175° C. to 400° C. under pressure. These patentsindicate that the synthetic method used therein was as set forth in U.S.Pat. No. 4,463,193, to Johnson. The Ford et al. '247 patent is also saidto be directed to color reduction of polyamines by reaction at elevatedtemperature and pressure in the presence of a hydrogenation catalyst anda hydrogen atmosphere. European Patent No. EP450709 to King et al. issaid to be directed to a process for the preparation oftriethylenetetramine and N-(2-aminoethyl)ethanolamine by condensation ofan alkylenamine and an alkylene glycol in the presence of a condensationcatalyst and a catalyst promoter at a temperature in excess of 260° C.

Russian Patent No. RU2186761, to Zagidullin, proposed synthesis ofdiethylenetriamine by reaction of dichloroethane with ethylenediamine.Ethylenediamine has previously been said to have been used in thesynthesis of N-carboxylic acid esters as reported in U.S. Pat. No.1,527,868, to Hartmann et al.

Japanese Patent No. 06065161 to Hara et al. is said to be directed tothe synthesis of polyethylenepolyamines by reacting ethylenediamine withethanolamine in the presence of silica-treated Nb205 supported on acarrier. Japanese Patent No. JP03047154 to Watanabe et al., is said tobe directed to production of noncyclic polyethylenepolyamines byreaction of ammonia with monoethanolamine and ethylenediamine.Production of non-cyclic polyethylenepolyamines by reaction ofethylenediamine and monoethanolamine in the presence of hydrogen or aphosphorous-containing substance was said to be reported in JapanesePatent No. JP03048644. Regenerative preparation of linearpolyethylenepolyamines using a phosphorous-bonded catalyst was proposedin European Patent No. EP115,138, to Larkin et al.

A process for preparation of alkyleneamines in the presence of a niobiumcatalyst was said to be provided in European Patent No. 256,516, toTsutsumi et al. U.S. Pat. No. 4,584,405, to Vanderpool, reported thecontinuous synthesis of essentially noncyclic polyethylenepolyamines byreaction of monoethanolamine with ethylenediamine in the presence of anactivated carbon catalyst under a pressure between about 500 to about3000 psig., and at a temperature of between about 200° C. to about 400°C. Templeton, et al., reported on the preparation of linearpolyethylenepolyamides asserted to result from reactions employingsilica-alumina catalysts in European Patent No. EP150,558.

Production of triethylenetetramine dihydrochloride was said to have beenreported in Kuhr et al., Czech Patent No. 197,093, via conversion oftriethylenetetramine to crystalline tetrahydrochloride and subsequentlyto triethylenetetramine dihydrochloride. “A study of efficientpreparation of triethylenetetramine dihydrochloride for the treatment ofWilson's disease and hygroscopicity of its capsule,” Fujito, et al.,Yakuzaigaku, 50:402-8 (1990), is also said to be directed to productionof triethylenetetramine.

Preparation of triethylenetetramine salts used for the treatment ofWilson's disease was said to be reported in “Treatment of Wilson'sDisease with Triethylene Tetramine Hydrochloride (Trientine),” Dubois,et al., J. Pediatric Gastro. & Nutrition, 10:77-81 (1990); “Preparationof Triethylenetetramine Dihydrochloride for the Treatment of Wilson'sDisease,” Dixon, et al., Lancet, 1(1775):853 (1972); “Determination ofTriethylenetetramine in Plasma of Patients by High-Performance LiquidChromatography,” Miyazaki, et al., Chem. Pharm. Bull., 38(4): 1035-1038(1990); “Preparation of and Clinical Experiences with Trien for theTreatment of Wilson's Disease in Absolute Intolerance ofD-penicillamine,” Harders, et al., Proc. Roy. Soc. Med., 70:10-12(1977); “Tetramine cupruretic agents: A comparison in dogs,” Allen, etal., Am. J. Vet. Res., 48(1):28-30 (1987); and “Potentiometric andSpectroscopic Study of the Equilibria in the AqueousCopper(II)-3,6-Diazaoctane-1,8-diamine System,” Laurie, et al., J.C.S.Dalton, 1882 (1976).

Preparation of triethylenetetramine salts by reaction of alcoholsolutions of amines and acids was said to be reported in Polish PatentNo. 105793, to Witek. Preparation of triethylenetetramine salts was alsoasserted in “Polycondensation of polyethylene polyamines with aliphaticdicarboxylic acids,” Witek, et al., Polimery, 20(3):118-119 (1975).

Baganz, H., and Peissker, H., Chem. Ber., 1957; 90:2944-2949; Haydock,D. B., and Mulholland, T. P. C., J. Chem. Soc., 1971; 2389-2395; andRehse, K., et al., Arch. Pharm., 1994; 393-398, report on Streckersyntheses. Use of Boc and other protecting groups has been described.See, for example, Spicer, J. A. et al., Bioorganic & MedicinalChemistry, 2002; 10: 19-29; Klenke, B. and Gilbert, I. H., J. Org.Chem., 2001; 66: 2480-2483.

Existing methods of synthesis of triethylenetetramines, andpolyethylenepolyamines, are unsatisfactory. For instance, they oftenrequire high temperature and pressure. A method for production of morepure triethylenetetramines at high yield under more favorable conditionsincluding, for example, at more manageable temperatures and pressures,would be desirable. Such methods have been invented and are describedand claimed herein.

BRIEF DESCRIPTION OF THE INVENTION

The inventions described and claimed herein have many attributes andencompass many embodiments including, but not limited to, those setforth in this Summary. The inventions described and claimed herein arenot limited to or by the features or embodiments identified in thisSummary, which is included for purposes of illustration only and notrestriction.

Methods for synthesis of triethylenetetramines are provided.Triethylenetetramines, and triethylenetetramine salts and polymorphs andcrystals thereof in high yields and purity are obtained.

In one embodiment at least one intermediate is synthesized that may becrystallized. Intermediates which may, but need not be, crystallizedinclude protected dinitrile intermediates. Crystallized dinitrileintermediates may be used in the synthesis of a triethylenetetramine.

In one embodiment, synthesis of a triethylenetetramine salt includessynthesis of a dinitrile from ethylenediamine and other startingmaterials, including, for instance, by alkylation by haloacetonitrile(for example, chloroacetonitrile or bromoacetonitrile). A Streckersynthesis using formaldehye and a cyanide salt may also be used to forma dinitrile from ethylenediamine. The resulting dinitrile is derivatized(optionally in situ) with a protecting group or groups, for example,benzaldehyde to form a benzaldehyde-protected dinitrile or Boc₂O(di-tert-butyl dicarbonate) to form a Boc-protected dinitrile. Theresulting protected dinitrile is optionally purified by crystallizationor another method. The protected dinitrile is reduced to form aprotected diamine, and the protected diamine is deprotected byhydrolysis with an acid to form a triethylenetetramine salt, including atriethylenetetramine primary salt, a triethylenetetramine secondarysalt, a triethylenetetramine tertiary salt and/or a triethylenetetraminequaternary salt. Optionally, further reaction of a triethylenetetraminetertiary salt or a triethylenetetramine quaternary salt with a base in asolvent, followed by reaction with a concentrated acid, may be used toform a triethylenetetramine secondary salt.

In another embodiment, synthesis of a triethylenetetramine salt includessynthesis of a dinitrile from ethylenediamine and other startingmaterials, including, for instance, by alkylation by haloacetonitrile(for example, chloroacetonitrile or bromoacetonitrile). A Streckersynthesis using formaldehye and a cyanide salt may also be used to forma dinitrile from ethylenediamine. The resulting dinitrile is thenderivatized (optionally in situ) with a protecting group or groups, forexample, benzaldehyde to form a benzaldehyde-protected dinitrile orBoc₂O (di-tert-butyl dicarbonate) to form a Boc-protected dinitrile. Theresulting protected dinitrile is optionally purified by crystallizationor another method. The protected dinitrile is reduced to form aprotected diamine. The diamine is derivatized with a protecting group orgroups to form a protected derivative, for example, with benzaldehyde toform a tri-benzaldehyde protected derivative. The protected derivativeis optionally purified by crystallization or another method. A furtherreaction includes deprotection of the protected derivative andpreparation of a triethylenetetramine salt, including atriethylenetetramine primary salt, a triethylenetetramine secondarysalt, a triethylenetetramine tertiary salt, and/or atriethylenetetramine quaternary salt. Optionally, further reaction of atriethylenetetramine tertiary salt or a triethylenetetramine quaternarysalt with a base in a solvent, followed by reaction with a concentratedacid, may be used to form a triethylenetetramine secondary salt.

In another embodiment the protected diamine undergoes a quenching stepto remove remaining reduction materials.

In still another embodiment a triethylenetetramine salt is prepared bythe addition of an acid, which optionally, has been dissolved in asolvent prior to its addition to the protected diamine. The subsequentcrystals formed are then re-crystallized in solution.

The invention also provides preparation of a purifiedtriethylenetetramine salt by derivatization of an impuretriethylenetetramine with a protecting group or protecting groups toform a protected derivative. For instance, an impuretriethylenetetramine may be derivatized with benzaldehyde, forming atribenzaldehyde protected derivative. The protected derivative ispurified by crystallization or another method. A further reactionincludes deprotection of the protected derivative and preparation of atriethylenetetramine salt, including a triethylenetetramine primarysalt, a triethylenetetramine secondary salt, a triethylenetetraminetertiary salt or a triethylenetetramine quaternary salt. Optionally,further reaction of a quaternary or tertiary triethylenetetramine saltwith a base in a solvent, followed by reaction with a concentrated acid,may be used to form a triethylenetetramine secondary salt.

Also provided are polymorphs of triethylenetetramine salts and methodsfor their preparation. These polymorphs include polymorphs oftriethylenetetramine disuccinate, triethylenetetraminetetrahydrochloride, and triethylenetetramine dihydrochloride.

In one embodiment a triethylenetetramine disuccinate polymorph has adifferential scanning calorimetry (DSC) extrapolated onset/peak meltingtemperature of from between about 170° C. to about 190° C.

In one embodiment a Form I polymorph of triethylenetetraminedihydrochloride has a DSC extrapolated onset/peak melting temperature offrom between about 111° C. to about 132° C.

In one embodiment a Form II polymorph of triethylenetetraminedihydrochloride is characterized by a DSC extrapolated onset/peakmelting temperature of from between about 170° C. to about 190° C.

In certain embodiments, crystalline triethylenetetramine and saltsthereof are provided. These include crystalline triethylenetetraminemaleate (e.g., triethylenetetramine tetramaleate andtriethylenetetramine tetramaleate dihydrate), triethylenetetraminefumarate (e.g., triethylenetetramine tetrafumarate andtriethylenetetramine tetrafumarate tetrahydrate), andtriethylenetetramine succinate (e.g, triethylenetetramine disuccinateanhydrate). Different triethylenetetramine crystals include thosecomprising the geometric structures, unit cell structures, andstructural coordinates set forth herein.

In certain embodiments, substantially pure triethylenetetraminedisuccinate is provided. A pharmaceutical composition comprisingsubstantially pure triethylenetetramine disuccinate is also provided.

In certain embodiments, substantially pure triethylenetetraminedisuccinate dihydrate is provided. In other embodiments, apharmaceutical composition comprising substantially puretriethylenetetramine disuccinate anhydrate is provided.

In other embodiments, substantially pure triethylenetetraminedisuccinate tetrahydrate is provided. In certain embodiments, apharmaceutical composition comprising substantially puretriethylenetetramine disuccinate tetrahydrate is also provided.

In certain embodiments, crystalline triethylenetetramine succinate inthe form of a crystal having alternating layers of triethylenetetraminemolecules and succinate molecules is provided. The triethylenetetraminesuccinate may be a triethylenetetramine disuccinate. Thetriethylenetetramine disuccinate may be a triethylenetetraminedisuccinate anhydrate. Pharmaceutical compositions comprisingcrystalline triethylenetetramine succinate in the form of a crystalhaving alternating layers of triethylenetetramine molecules andsuccinate molecules are also provided, including pharmaceuticalcompositions having crystalline triethylenetetramine disuccinateanhydrate in the form of a crystal having alternating layers oftriethylenetetramine molecules and succinate molecules.

In certain embodiments, substantially pure triethylenetetramine maleateis provided. A pharmaceutical composition comprising substantially puretriethylenetetramine maleate is also provided.

In certain embodiments, substantially pure triethylenetetraminetetramaleate is provided. In other embodiments, a pharmaceuticalcomposition comprising substantially pure triethylenetetraminetetramaleate is provided.

In other embodiments, substantially pure triethylenetetraminetetramaleate dihydrate is provided. In certain embodiments, apharmaceutical composition comprising substantially puretriethylenetetramine tetramaleate anhydrate is also provided.

In certain embodiments, a composition comprising crystallinetriethylenetetramine maleate in the form of a crystal having alternatinglayers of triethylenetetramine molecules and maleate molecules isprovided. The triethylenetetramine maleate may be a triethylenetetraminetetramaleate. The triethylenetetramine maleate may be atriethylenetetramine tetramaleate dihydrate. Pharmaceutical compositionscomprising crystalline triethylenetetramine maleate in the form of acrystal having alternating layers of triethylenetetramine molecules andmaleate molecules are also provided, including pharmaceuticalcompositions having crystalline triethylenetetramine tetramaleatedihydrate in the form of a crystal having alternating layers oftriethylenetetramine molecules and maleate molecules.

In certain embodiments, substantially pure triethylenetetramine fumarateis provided. A pharmaceutical composition comprising substantially puretriethylenetetramine fumarate and/or a pharmaceutically acceptableexcipient is also provided.

In certain embodiments, substantially pure triethylenetetraminetetrafumarate is provided. In other embodiments, a pharmaceuticalcomposition comprising substantially pure triethylenetetraminetetrafumarate is provided.

In other embodiments, substantially pure triethylenetetraminetetrafumarate tetrahydrate is provided. In certain embodiments, apharmaceutical composition comprising substantially puretriethylenetetramine tetrafumarate tetrahydrate is provided.

In certain embodiments, a composition comprising crystallinetriethylenetetramine fumarate in the form of a crystal havingalternating layers of triethylenetetramine molecules and fumaratemolecules is provided. The triethylenetetramine fumarate may be atriethylenetetramine tetrafumarate. The triethylenetetramine fumaratemay be a triethylenetetramine tetrafumarate tetrahydrate. Pharmaceuticalcompositions comprising crystalline triethylenetetramine fumarate in theform of a crystal having alternating layers of triethylenetetraminemolecules and fumarate molecules are also provided, includingpharmaceutical compositions having crystalline triethylenetetraminetetrafumarate tetrahydrate in the form of a crystal having alternatinglayers of triethylenetetramine molecules and fumarate molecules.

In certain embodiments, a crystal of a triethylenetetramine disuccinatehaving a C 2/c (no. 15) space group with measured unit cell dimensionsof a=14.059(5) Å, b=9.169(5) Å, c=13.647(5) Å, and 0=92.47(0) Å, isprovided. Cell volume is 1757.56(130) Å³.

In certain embodiments, A crystal of a triethylenetetramine tetramaleatehaving a P 1 2/c 1 (no. 13) space group with measured unit celldimensions of a=13.261(5) Å, b=9.342 Å, c=11.266 Å, and 3=91.01(0) Å, isprovided. Cell volume is 1395.46(110) Å³.

In certain embodiments, a crystal of a triethylenetetraminetetrafumarate having a P m n a (no. 53) space group with measured unitcell dimensions of a=13.9031(3) Å, b=7.9589(2) Å, c=14.6554(3) Å, and(3=90 Å, is provided. Cell volume is 1621.67(6) Å³.

In certain embodiments, a crystal of a triethylenetetramine disuccinatehaving the structure defined by the co-ordinates of Tables 1A-1C isprovided.

In certain embodiments, a crystal of a triethylenetetramine tetramaleatedihydrate having the structure defined by the co-ordinates of Tables2A-2C is provided.

In certain embodiments, a crystal of a triethylenetetraminetetrafumarate tetrahydrate having the structure defined by theco-ordinates of Tables 3A-3C is provided.

Also provided are pharmaceutical compositions, including pharmaceuticalcompositions comprising substantially pure triethylenetetraminedisuccinate. triethylenetetramine disuccinate anhydrate,triethylenetetramine tetramaleate, triethylenetetramine tetramaleatedihydrate, triethylenetetramine tetrafumarate, or triethylenetetraminetetrafumarate tetrahydrate, that also include one or morepharmaceutically acceptable excipients, carriers, and/or additivessuitable for oral or parenteral application.

Uses of the disclosed and claimed compounds and pharmaceuticalcompositions in the treatment of various diseases, disorders andconditions are also provided.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a reaction scheme according to one embodiment of theinvention for synthesizing a triethylenetetramine salt.

FIG. 2 shows a reaction scheme according to another embodiment of theinvention for synthesizing a triethylenetetramine salt.

FIG. 3 shows a reaction scheme according to a further embodiment of theinvention for synthesizing a triethylenetetramine salt.

FIG. 4 shows an ¹H-NMR spectrum of benzaldehyde-protected dinitrile(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile as synthesizedin Example 1. NMR values include a frequency of 400.13 Mhz, a 1Hnucleus, number of transients is 16, points count of 32768, pulsesequence of zg30, and sweep width of 8278.15 Hz.

FIG. 5 shows an ¹H-NMR spectrum of the tri-benzaldehyde derivativebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminein CDCl₃, as synthesized in Example 2. NMR values include a frequency of400.13 Mhz, a 1H nucleus, number of transients is 16, points count of32768, pulse sequence of zg30, and sweep width of 8278.15 Hz.

FIG. 6 shows an ¹H-NMR spectrum of a triethylenetetramine hydrochloridesalt in D₂O, as synthesized in Example 3. NMR values include a frequencyof 400.13 Mhz, a 1H nucleus, number of transients is 16, points count of32768, pulse sequence of zg30, and sweep width of 8278.15 Hz.

FIG. 7 shows an infrared spectrum of a polymorph of triethylenetetraminedisuccinate.

FIG. 8 shows a DSC graph of a polymorph of triethylenetetraminedisuccinate.

FIG. 9 shows an infrared spectrum of the Form I polymorph oftriethylenetetramine dihydrochloride.

FIG. 10 shows a DSC graph of the Form I polymorph oftriethylenetetramine dihydrochloride.

FIG. 11 shows an infrared spectrum of the Form II polymorph oftriethylenetetramine dihydrochloride.

FIG. 12 shows a DSC graph of the Form II polymorph oftriethylenetetramine dihydrochloride.

FIG. 13 shows a 2×2×2 unit cell of triethylenetetramine disuccinate witha view along (I) [010], (II) [100] and (III) [001].

FIG. 14 shows a coordination sphere for (I) triethylenetetramine and(II) triethylenetetramine disuccinate.

FIG. 15 shows an X-ray powder pattern of the triethylenetetraminedisuccinate powder material in comparison with the calculated powderpattern obtained from the single crystal structure data fortriethylenetetramine disuccinate.

FIG. 16 shows an X-ray powder pattern of the re-crystallisedtriethylenetetramine disuccinate powder material in comparison with thecalculated powder pattern obtained from the single crystal structuredata for triethylenetetramine disuccinate.

FIG. 17 shows (I) a unit cell of triethylenetetramine tetramaleate and(II) 2×2×2 unit cells of triethylenetetramine tetramaleate with viewalong [010], and (III) 2×2×2 unit cells of triethylenetetraminetetramaleate with view along [100].

FIG. 18 shows the coordination sphere of triethylenetetraminetetramaleate with view along (I) [001] and (II) [110].

FIG. 19 shows the X-ray powder pattern of the triethylenetetraminetetramaleate powder in comparison with the calculated powder patternobtained from the single crystal structure data for triethylenetetraminetetramaleate.2H₂O.

FIG. 20 shows 2×2×2 unit cells of triethylenetetramine tetrafumarate,with view along (I) [010], (II) [001] and (III) [100].

FIG. 21 shows a coordination sphere of triethylenetetramine with viewalong (I) [100] and (II) [010].

FIG. 22 shows an X-ray powder pattern of triethylenetetraminetetrafumarate powder material in comparison with the calculated powderpattern obtained from the single crystal structure data fortriethylenetetramine tetrafumarate.4H₂O.

DETAILED DESCRIPTION OF THE INVENTION

The subject matter of this patent is also illustrated with reference tothe figures, including accompanying FIG. 1. FIG. 1 shows a summary of apreferred reaction scheme for the preparation of triethylenetetraminesalts based generally on the Strecker-synthesis of a dinitrile followedby reduction of the dinitrile. The reaction includes formation of adinitrile from ethylenediamine using formaldehyde and a cyanide saltunder acidic conditions. The resulting dinitrile is derivitized to forma benzaldehyde-protected dinitrile. The protected dinitrile is reducedto form a protected diamine, and the diamine is derivatized withbenzaldehyde to form a benzaldehyde protected derivative. The protectedderivative is deprotected by hydrolysis with an acid to form atriethylenetetramine salt, which may include a triethylenetetramineprimary salt, a triethylenetetramine secondary salt, atriethylenetetramine tertiary salt, or a triethylenetetramine quaternarysalt. Other reaction schemes, including reaction schemes such as thoseshown in FIGS. 2 and 3, are also provided.

FIG. 2 shows a reaction scheme, similar to that of FIG. 1, wherein theprotected diamine is directly hydrolyzed to form a triethylenetetraminesalt, which may be a triethylenetetramine primary salt, atriethylenetetramine secondary salt, a triethylenetetramine tertiarysalt, or a triethylenetetramine quaternary salt.

FIG. 3 shows another reaction scheme for the preparation oftriethylenetetramine salts based generally on the Strecker-synthesis ofa dinitrile, wherein dinitrile is derivatized with Boc₂O (di-tert-butyldicarbonate) to form a Boc-protected dinitrile, the Boc-protecteddinitrile is reduced in an aqueous solution of ethanol and ammonia inthe presence of Raney-nickel and dihydrogen to form a protected diamine,and the protected diamine is purified by precipitation in isopropanol.The resulting compound is deprotected and hydrolyzed with an acid toform a triethylenetetramine salt, which may include atriethylenetetramine primary salt, a triethylenetetramine secondarysalt, a triethylenetetramine tertiary salt, or a triethylenetetraminequaternary salt.

The patented invention includes individual steps, reaction conditions,and reactants and chemicals, as well as routes for synthesis oftriethylenetetramines in high yield and purity, including by reductionof a protected dinitrile or by reduction of other intermediates orstarting materials.

The patented inventions also include certain compounds that haveutility, for example, as intermediates for synthesis oftriethylenetetramine. Intermediates may be independently isolated andpurified and/or crystallized, including during and as a part of themethods of synthesis provided herein. Isolated and purified and/orcrystallized intermediates may also be stored for later use.

The steps and routes of synthesis are effective for preparation of avariety of triethylenetetramine salts. Such salts include, for example,triethylenetetramine succinate, triethylenetetramine diuccinate,triethylenetetramine diuccinate anhydrate, triethylenetetramine maleate,triethylenetetramine tetramaleate, triethylenetetramine tetramaleatedihydrate, triethylenetetramine fumarate, triethylenetetraminetetrafumarate, triethylenetetramine tetrafumarate tetrahydrate.

Also provided are polymorphs of various triethylenetetramine salts andmethods for their production. In one embodiment a triethylenetetraminedisuccinate polymorph has a differential scanning calorimetry (DSC)extrapolated onset/peak melting temperature of from between about 170°C. to about 190° C. See FIGS. 7 and 8.

In another embodiment a Form I polymorph of triethylenetetraminedihydrochloride has a DSC extrapolated onset/peak melting temperature offrom between about 111° C. to about 132° C. See FIGS. 9 and 10. In yetanother embodiment a Form II polymorph of triethylenetetraminedihydrochloride is characterized by a DSC extrapolated onset/peakmelting temperature of from between about 170° C. to about 190° C. SeeFIGS. 11 and 12.

Also provided are triethylenetetramine salts of high purity, methods fortheir preparation, and dosage forms including triethylenetetraminesalts.

Especially preferred individual compounds of the invention aretriethylenetetramine succinate, triethylenetetramine disuccinate, andtriethylenetetramine disuccinate hydrates.

Triethylenetetramine pharmaceutical compositions including one or moreof, for example, triethylenetetramine succinate, triethylenetetraminedisuccinate, triethylenetetramine disuccinate hydrate,triethylenetetramine maleate, triethylenetetramine tetramaleate,triethylenetetramine tetramaleate dihydrate, triethylenetetraminefumarate, triethylenetetramine tetrafumarate, triethylenetetraminetetrafumarate tetrahydrate are also provided, as are methods for theiruse. The pharmaceutical compositions may include, for example, one ormore pharmaceutically acceptable excipients, carriers, and/or additivessuitable for oral or parenteral administration.

The product formed by the described processes is substantially pure,that is, substantially free from any other compounds. Preferably, itcontains less than 10% impurities, and more preferably, less than about5% impurities, and even more preferably, less than about 1% impurities.The product thus formed is also preferably substantially pure, i.e.,contains less than 10% impurity, more preferably less than 5% impurity,and still more preferably less than 1% impurity. The present inventionalso includes a substantially pure anhydrous crystalline form oftriethylenetetramine disuccinate. The term “substantially pure” meansthat a sample of the relevant anhydrous crystalline form oftriethylenetetramine disuccinate contains more than 90% of a singlepolymorphic form, preferably more than 95% of a single polymorphic form,and still more preferably more than 99% of a single polymorphic form.The present invention also provides substantially pure crystalline formsof triethylenetetramine tetramaleate dihydrate and triethylenetetraminetetrafumarate tetrahydrate, as described herein.

The term “amine protecting group” as used herein refers to any moietythat is used to protect at least one —NH— moiety and/or at least one—NH₂ moiety by replacement of hydrogen. Any moiety that is, for example,relatively inert to reaction conditions under which a nitrile is reducedmay be used. The resulting protected structure may be linear or cyclic,and may include one or more amine protecting groups. Examples of amineprotecting groups useful in the present invention include, by way ofexample only, methyl carbamate, ethyl carbamate, benzyl carbamate,tert-butyl carbamate, tert-butyloxycarbonyl (Boc), cyclohexanone,2,2,6,6-tetramethyl cyclohexanone, anthrone, an alkyl group, an arylgroup, or an aromatic alkyl group. Other amine protecting groupssuitable for use in the invention are described, for example, inProtective Groups in Organic Synthesis, Third Edition, T. W. Green andP. G. M. Wuts (Wiley-Interscience, 1999). Others will be known to thosein the art.

The term “alkyl” includes saturated aliphatic groups, includingstraight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, nonyl, decyl, etc.), branched-chain alkyl groups(isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl (alicyclic) groups(cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkylsubstituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.The term alkyl further includes alkyl groups, which comprise oxygen,nitrogen, sulfur or phosphorous atoms replacing one or more carbons ofthe hydrocarbon backbone. The term “aromatic-alkyl” includes alkylgroups substituted with one or more aryl groups.

The term “aryl” includes groups with aromaticity, including 5- and6-membered single-ring aromatic groups that may include from zero tofour heteroatoms as well as multicyclic systems with at least onearomatic ring. Examples of aryl groups include benzene, phenyl, pyrrole,furan, thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole,pyrazole, oxazole, isooxazole, pyridine, pyrazine, pyridazine, andpyrimidine, and the like. Furthermore, the term “aryl” includesmulticyclic aryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene,benzoxazole, benzodioxazole, benzothiazole, benzoimidazole,benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline,napthridine, indole, benzofuran, purine, benzofuran, deazapurine, orindolizine. Those aryl groups having heteroatoms in the ring structuremay also be referred to as “aryl heterocycles”, “heterocycles,”“heteroaryls” or “heteroaromatics”. The aromatic ring can be substitutedat one or more ring positions with such substituents as described above,as for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl,alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Aryl groups can also be fused or bridged withalicyclic or heterocyclic rings which are not aromatic so as to form amulticyclic system (e.g., tetralin, methylenedioxyphenyl).

The term “imidazolidine derivative-forming amine protecting groupreagent” as used herein refers to a reactant used to protect more thanone —NH— moiety by formation of an imidazolidine derivative. Examples ofimidazolidine derivative-forming amine protecting group reagentsinclude, for example, but are not limited to, an aldehyde, a ketone,formaldehyde, a substituted aromatic aldehyde, a substituted aliphaticaldehyde, a substituted alkyl-aromatic aldehyde, a substituted aromaticketone, a substituted aliphatic ketone, and a substituted alkyl-aromaticketone.

The term “amine protecting group reagent” as used herein refers to areactant used to protect at least one —NH— moiety or at least one —NH₂moiety including imidazolidine derivative-forming amine protecting groupreagents. For example, the reagent di-tert-butyl dicarbonate (Boc₂O) maybe used to protect about two equivalents of —NH— moiety for every oneequivalent of Boc₂O used in a reaction. Amine protecting group reagentsinclude, for example, Boc₂O, an aldehyde, a ketone, formaldehyde, asubstituted aromatic aldehyde, a substituted aliphatic aldehyde, asubstituted alkyl-aromatic aldehyde, a substituted aromatic ketone, asubstituted aliphatic ketone, and a substituted alkyl-aromatic ketone.

The term “treating”, “treat” or “treatment” as used herein includespreventative (e.g., prophylactic) and palliative treatment.

By “pharmaceutically acceptable” is meant the carrier, diluent,excipients, and/or salt must be compatible with the other ingredients ofthe formulation, and not deleterious to the recipient thereof.

In one embodiment, synthesis of a triethylenetetramine includessynthesis of a dinitrile from ethylenediamine and other startingmaterials, including, for instance, by alkylation using anhaloacetonitrile (for example, chloroacetonitrile or bromoacetonitrile).A Strecker synthesis using formaldehye and a cyanide salt, for example,KCN, may be used to form a dinitrile from ethylenediamine. The resultingdinitrile is derivatized (optionally in situ) with a protecting group orgroups. For example, the dinitrile may be derivatized with benzaldehydeto form a benzaldehyde-protected dinitrile or with Boc₂O (di-tert-butyldicarbonate) to form a Boc-protected dinitrile. The resulting protecteddinitrile is optionally purified by crystallization or another method.The protected dinitrile is reduced to form a protected diamine, and theprotected diamine is deprotected by hydrolysis with an acid to form atriethylenetetramine salt, for example, a triethylenetetramine primarysalt, a triethylenetetramine secondary salt, a triethylenetetraminetertiary salt or a triethylenetetramine quaternary salt. Optionally,further reaction of a triethylenetetramine tertiary salt or atriethylenetetramine quaternary salt with a base in a solvent, followedby reaction with a concentrated acid, may be used to form atriethylenetetramine secondary salt.

In another embodiment, synthesis of a triethylenetetramine salt includessynthesis of a dinitrile from ethylenediamine and other startingmaterials, including, for instance, by alkylation by haloacetonitrile(for example, chloroacetonitrile or bromoacetonitrile). A Streckersynthesis using formaldehye and a cyanide salt may also be used to forma dinitrile from ethylenediamine. The resulting dinitrile is thederivatized (optionally in situ) with a protecting group or groups, forexample, benzaldehyde to form a benzaldehyde-protected dinitrile orBoc₂O (di-tert-butyl dicarbonate) to form a Boc-protected dinitrile. Theresulting protected dinitrile is optionally purified by crystallizationor another method. The protected dinitrile is reduced to form aprotected diamine. The diamine is derivatized with a protecting group orgroups to form a protected derivative, for example, with benzaldehyde toform a tri-benzaldehyde protected derivative. The protected derivativeis optionally purified by crystallization or another method. A furtherreaction includes deprotection of the protected derivative andpreparation of a triethylenetetramine salt, including atriethylenetetramine primary salt, a triethylenetetramine tertiary salt,a triethylenetetramine secondary salt, or a triethylenetetraminequaternary salt. Optionally, further reaction of a triethylenetetraminetertiary salt or a triethylenetetramine quaternary salt with a base in asolvent, followed by reaction with a concentrated acid, may be used toform a triethylenetetramine secondary salt.

In another embodiment the protected diamine undergoes a quenching stepto remove remaining reduction materials.

In another embodiment a triethylenetetramine salt is prepared by theaddition of an acid, which optionally, has been dissolved in a solventprior to its addition to the protected diamine. The subsequent crystalsformed are then re-crystallized in solution.

The invention also provides preparation of a purifiedtriethylenetetramine salt by derivatization of an impuretriethylenetetramine with a protecting group or protecting groups toform a protected derivative. For instance, an impuretriethylenetetramine may be derivatized with benzaldehyde, forming atribenzaldehyde protected derivative. The protected derivative ispurified by crystallization or another method. A further reactionincludes deprotection of the protected derivative and preparation of atriethylenetetramine salt, including a triethylenetetramine primarysalt, a triethylenetetramine secondary salt, a triethylenetetraminetertiary salt or a triethylenetetramine quaternary salt. Optionally,further reaction of a quaternary or tertiary triethylenetetramine saltwith a base in a solvent, followed by reaction with a concentrated acid,may be used to form a triethylenetetramine secondary salt.

A. Preparation of a Dinitrile

In one embodiment, a dinitrile intermediate such as that shown as (V) ora derivative thereof is formed.

The dinitrile intermediate may be formed by a variety of routes, from anumber of reagents, and under a variety of conditions. Routes, reagentsand conditions include those set forth in more detail herein, and in thebelow Examples.

1. Dinitrile Preparation by Alkylation

One method for forming a dinitrile intermediate is by alkylation of astarting compound. In one route, a dinitrile intermediate is formed byalkylation of a diamine with a haloacetonitrile in the presence of abase and in a solvent. Suitable diamines for use as starting materialfor the preparation of triethylenetetramine include but are not limitedto, for example, ethylenediamine, ethylenediamine hydrochloride salt,ethylenediamine hydrobromide salt, ethylenediamine diacetate salt, orother salts of ethylenediamine. Other suitable starting materialsinclude those materials that may be changed to ethylenediamine in situ,including but not limited to ethene-1,2-diamine.

A number of haloacetonitriles are suitable for use in the alkylation,including but not limited to, for example, chloroacetonitrile andbromoacetonitrile. Although at least about two equivalents ofhaloacetonitrile are generally necessary for full reaction of eachequivalent of ethylenediamine, the reaction may be conducted with lesshaloacetonitrile if incomplete conversion is desired. Greater than abouttwo equivalents of haloacetonitrile may be used to increase the rate ofthe reaction. Use of greater than about three equivalents ofhaloacetonitrile is less favored as it may result in overalkylation ofthe expected dinitrile.

Bases suitable for use in alkylation include but are not limited to, forexample, potassium carbonate, sodium carbonate, sodium hydroxide,potassium hydroxide, and sodium tert-butylate. The amount of base addedto the reaction may vary between about 0.1 equivalents and about 2.0equivalents of diamine starting material; the more soluble the base isin the selected solvent, the less base need be used. For example, in oneembodiment of the invention, the solvent used is acetonitrile and about2 equivalents of potassium carbonate are used; potassium carbonate issparingly soluble in acetonitrile.

The alkylation may be performed in a number of solvents, including butnot limited to acetonitrile, tetrahydrofuran (also referred to herein as“THF”), an ether, a hydrocarbon such as an alkane, or mixtures thereof.Preferably the solvent used in the reaction is non reactive at thetemperature at which the reaction is conducted. The reaction may beconducted at a temperature from about the melting point of the selectedsolvent to about 25° C. Lower temperatures will generally lead to aslower rate of reaction.

In one embodiment of the invention, alkylation is performed by adding asolution of about 2 equivalents of chloroacetonitrile in acetonitrile toa mixture of about 1 equivalent of ethylenediamine and about 2equivalents of potassium carbonate in acetonitrile. The addition occursover about 30 minutes at a temperature of about 25° C., leading tocompletion of the reaction in about 21 hours and formation of adinitrile intermediate.

2. Dinitrile Preparation with a Formaldehyde Starting Material

Another route for formation of a dinitrile intermediate such as (V), ora derivative thereof, is through reaction of formaldehyde with otherstarting materials. In one route, a dinitrile intermediate is formed byreaction of formaldehyde with an inorganic cyanide salt, anethylenediamine source, and an acid in a solvent.

Suitable ethylenediamine sources include but are not limited toethylenediamine, ethylenediamine dihydrochloride salt, ethylenediaminedihydrobromide salt, ethylenediamine diacetate salt, or other salts ofethylenediamine. Other suitable starting materials include materialsthat are reacted to form ethylenediamine in situ, including but notlimited to ethene-1,2-diamine. Ethylenediamine is preferred.

Acids suitable for use in the reaction include but are not limited tohydrochloric acid hydrobromic acid, and phosphoric acid, though an acidneed not be added to the reaction if an ethylenediamine salt is used asa starting material. About 2 equivalents of acid are generally used forevery equivalent of ethylenediamine starting material. Fewer equivalentsof acid may be used, including between about 1 and about 2 equivalentsof acid, though use of fewer than about two equivalents of acid islikely to decrease the yield of the dinitrile intermediate. Betweenabout 2 and about 3 equivalents of acid may be used, although use ofexcess acid is not preferred due to the possibility for the reaction toliberate hydrogen cyanide gas and this liberation will reduce the amountof cyanide available to produce the dinitrile intermediate. Liberationof hydrogen cyanide gas requires that precautions be taken due to thetoxicity of hydrogen cyanide. If an ethylenediamine salt in solution isused as a starting material, 2 equivalents of acid for every 1equivalent of ethylenediamine are generally preferred. If an acid isused, hydrochloric acid is preferred.

The formaldehyde is, for example, formaldehyde in aqueous solution orformaldehyde trimer in paraffin. Formaldehyde may be present in anamount between about 1.8 equivalents for each equivalent ofethylenediamine to about 3.0 equivalents for each equivalent ofethylenediamine. In a preferred embodiment of the invention, about 2.2equivalents of formaldehyde are used for every equivalent ofethylenediamine.

Cyanide salts used to practice the invention include but are not limitedto inorganic cyanide salts such as sodium cyanide, potassium cyanide,lithium cyanide, magnesium cyanide, and hydrogen cyanide. Use ofhydrogen cyanide, a gas, is not preferred due to toxicity and therequirement that the reaction be conducted under pressure. Heavy metalcyanides such as silver cyanide, gold cyanide and copper cyanide mayalso be used but are not preferred due to the tendency of complexationwith the dinitrile product. About two equivalents of cyanide salt areused in the reaction for every equivalent of ethylenediamine. Fewerequivalents of cyanide salt may be used, for instance between about 1equivalents and about 2 equivalents, but use of fewer than about 2equivalents of cyanide salt will decrease yield of the dinitrile.Cyanide salts may also be used in amounts greater than about twoequivalents for each equivalent of ethylenediamine. In one preferredembodiment, 1.98 eq of cyanide, e.g., KCN, is used for the reaction.

The reaction may be conducted over a range of temperatures, includingbut not limited to temperatures between about −5° C. and about 35° C.,including, for example, from about 4° C. and about 25° C. In one morepreferred embodiment of the invention the reaction is conducted at about20° C. The reaction may be conducted at, but is not limited to beingconducted at, a pH in the range of from about 8 to about 14, from about9 to about 12, or from about 10 to about 11. After the complete additionof formaldehyde the pH may be adjusted to the ranges of from about 1 toabout 8, from about 2 to about 7, from about 3 to about 7, from about 4to about 7, from about 5 to about 6, or from about 6 to about 7. In apreferred embodiment of the invention, the temperature is from about 4°C. and about 25° C. and the pH is from about 9 to about 12.

A variety of solvents may be used, including water and other misciblenon reactive solvents that do not alter the reaction pH beyond anoperative range. The pH can be adjusted with any suitable buffer,including but not limited to, for example, acetic acid, any aceticbuffer or a phosphate buffer.

In one embodiment a dinitrile intermediate as shown in (V) is preparedby adding about two equivalents of potassium cyanide to about oneequivalent of ethylenediamine hydrochloride salt in water. A solution ofabout 2.2 equivalents of formaldehyde in water is added over about 75minutes, with the pH of the solution adjusted to about 5 with aceticacid following addition of the formaldehyde. A reaction to produce adinitrile intermediate is complete after about 17 hours at about 20° C.

In one embodiment, a solution of 1 equivalent of ethylenediamine HCl and1.98 equivalents of KCN in water are treated with 1.98 equivalents offormaldehyde to form the dinitrile intermediate as shown in (V). Theintermediate may be isolated by the addition of benzaldehyde at a pH offrom about 6 to about 7 in a two-phase system (water/n-butanol).

In a preferred embodiment, a solution of 1 equivalent of ethylenediamineand 1.98 equivalents of HCl are combined, together with 1.98 equivalentsof formaldehyde. A pH range of about 7.0 is used for the reaction. Inone preferred embodiment, a slight excess of ethylene diamine startingmaterial is used to achieve the desired pH range. Preferably, theformaldehyde is added last to enhance yield, and the formaldehyde isadded over 45 minutes followed by a reaction time of 2.5 hours at 17-20°C. for complete conversion of the Strecker reaction.

B. Protection of Secondary Amines and Isolation of ProtectedIntermediate

In another embodiment of the invention, a dinitrile as shown in (V), ora derivative thereof, is reacted with a suitable amine protecting groupreagent to protect secondary amines of the dinitrile as tertiary amines.The dinitrile may be formed as described above, for example, and may butneed not be first isolated.

Reaction with an amine protecting group reagent forms a protecteddinitrile. Protection of the secondary amines will prevent unfavorableoxidation or reduction of amine groups of the protected dinitrile duringsubsequent reactions, including reduction of the nitrile groups.Optionally, a protected dinitrile is purified by crystallization priorto further reaction.

1. Protection by Formation of an Acyclic Intermediate

In one embodiment of the invention, a dinitrile such as (V) is protectedby reaction with a suitable amine protecting group reagent to form aprotected dinitrile as shown in Formula (VI),

where the compound formed is a[{2-[cyanomethyl-R-amino]-ethyl}-R-amino]-acetonitrile, in which R is anamine protecting group. R may include, for example, but is not limitedto, methyl carbamate, ethyl carbamate, benzyl carbamate, tert-butylcarbamate, tert-butyloxycarbonyl (Boc), cyclohexanone,2,2,6,6-tetramethyl cyclohexanone, anthrone, an alkyl group, an arylgroup, or an aromatic alkyl group. Other suitable amine protectinggroups are described, for example, in Protective Groups in OrganicSynthesis, Third Edition, T. W. Green and P. G. M. Wuts(Wiley-Interscience, 1999), or otherwise known in the art.

A dinitrile intermediate, such as (V), may be reacted with a number ofamine protecting group reagents to affix the desired protecting groups.The dinitrile intermediate may be reacted with, for example,di-tert-butyl dicarbonate, tert-butyl carbonate, an aldehyde, a ketone,formaldehyde, a substituted aromatic aldehyde, a substituted aliphaticaldehyde, a substituted alkyl-aromatic aldehyde, a substituted aromaticketone, a substituted aliphatic ketone, or a substituted alkyl-aromaticketone. For complete reaction to occur, at least about two equivalentsof an amine protecting group should be available in the reaction. Wheretwo equivalents of an amine protecting group are liberated from oneequivalent of an amine protecting group reagent (for example, when Boc₂Ois used), only one equivalent of the amine protecting group reagent needbe used. Amine protecting group reagent(s) may be added to excess.

The reaction is conducted in at least one of a variety of solvents,including but not limited to water and acetonitrile. Solvent selectionwill be based on the identity of the protecting group. Preferably thesolvent used in the addition of the protecting group is the same as thatused in the formation of the dinitrile intermediate, which favorsconducting reactions serially and in the same vessel. The reaction isgenerally performed at temperatures between about −5° C. to about 35°C., for example, from about 4° C. to about 25° C.

The protected dinitrile is optionally purified and crystallized prior tosubsequent use or reaction. Purification and crystallization may beperformed by extraction, crystallization, or extraction andcrystallization from a solvent. Suitable solvents include, for example,ethers, alkanes including methylcyclohexane, ethyl acetate, or a mixtureof solvents including a mixture of ethyl acetate and methylcyclohexanein a ratio of about 3 parts ethyl acetate to about 10 partsmethylcyclohexane.

In one embodiment, an acyclic protected dinitrile is prepared byreacting a dinitrile such as (V) with about 1.2 equivalents of Boc₂O inacetonitrile. Complete conversion to a Boc-protected dinitrile (FormulaVI, where R is Boc) is achieved after about one hour at about 20° C.Purification and crystallization give a Boc-protected dinitrile ingreater than 80% yield.

2. Protection by Formation of a Dinitrile Including an ImidazolidineDerivative

In another embodiment of the invention, a dinitrile such as (V) isprotected by reaction with a suitable imidazolidine derivative-formingamine protecting group reagent to form a protected dinitrile as shown inFormula (IV), where the compound formed is(3-cyanomethyl-2-R₁-2-R₂-imidazolidin-1-yl)-acetonitrile, and where R₁and R₂ may be the same or different, and may be, for example, hydrogen,an alkyl group including from one to twelve carbon atoms, an aryl group,or an aromatic alkyl group.

The protected dinitrile of Formula (IV) may be formed by reaction of adinitrile with between about 0.9 equivalents and about two equivalentsof an imidazolidine derivative-forming amine protecting group reagent,including, for example, an aldehyde, a ketone, formaldehyde, asubstituted aromatic aldehyde, a substituted aliphatic aldehyde, asubstituted alkyl-aromatic aldehyde, a substituted aromatic ketone, asubstituted aliphatic ketone, or a substituted alkyl-aromatic ketone.The reaction is conducted at a pH in the range of from about 4 to about8, from about 6 to about 8, or from about 7 to about 8. A buffer isoptionally included to maintain a desired pH. The preferred pH is about7. Suitable buffers include but are not limited to sodium dihydrogenphosphate.

The reaction is conducted in a least one of a variety of solvents, whichinclude, for example, water or acetonitrile. Solvent selection is basedon the identity of the imidazolidine derivative-forming amine protectinggroup reagent. Preferably the solvent used is the same as that used inthe formation of the dinitrile, so that the reactions may be conductedserially and in the same vessel. The reaction is generally performed attemperatures between about −5° C. and about 35° C., or from about 4° C.to about 25° C.

Following synthesis of a protected dinitrile, the protected dinitrile isoptionally extracted with a solvent, including for example, ethylacetate, and optionally crystallized from a mixture of butanol andcyclohexane, for example. In another embodiment a protected dinitrile isremoved from solution by precipitation after addition of an alcohol.Suitable alcohols include, for example, but are not limited to,isopropanol, n-butanol, and t-butanol.

In one embodiment, a dinitrile is reacted with about 1 equivalent ofbenzaldehyde in water and stirred at about 20° C. for about 2 hours.Purification by crystallization in cyclohexane gives the protecteddinitrile in about 40% yield of crystalline product.

In another embodiment, a dinitrile is reacted with about 1.1 equivalentsof benzaldehyde in water and in the presence of a phosphate buffer. Thereaction is conducted at a pH of from about 6 to about 7. An excess ofbutanol is added to the reaction mixture to produce a biphasic mixture.The protected dinitrile derivative (I) precipitates in about two hours,resulting in about 76% to about 78% yield of crystalline product with apurity greater than about 99 area %. The protected dinitrile derivative(I) may be isolated and crystallized. In a preferred embodiment, thecrystallization step is carried out for at least 6 hours at 0° C.

In another embodiment, an intermediate for the production oftriethylenetetramines and salts thereof is2-(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetamide (shown in (VII),below) or 2-(3-carbamoylmethyl-2-phenyl-imidazolidin-1-yl)-acetamide(shown in (VIII), below), both of which may be reduced and converted totriethylenetetramine as set forth herein for (I).

C. Reduction of Nitrile Groups to Form Protected Diamine Intermediate

In a further step, a protected dinitrile is reacted with a reducingagent to create a protected diamine. As discussed further in Section D,below, the protected diamine is then either deprotected by hydrolysis inthe presence of an acid to form a triethylenetetramine salt (as shownand discussed in Section D(1)), for example, or the protected diamine isreacted with further protecting groups and purified by crystallization,then hydrolyzed to form a triethylenetetramine salt (as discussed inSection D(2)).

To form a protected diamine, a protected dinitrile is reduced with areducing agent in a solvent. Reducing agents that may be used include,for example, LiAlH₄ (also referred to herein as “LAH”), nickel-aluminumcatalyst/H₂, Raney-Nickel/H₂, NaAlH₄, Li(MeO)₃AlH, di-isobutyl aluminumhydride, sodium bis(2-methoxyethoxy)aluminum hydride, charcoalcatalyst/H₂ and platinum catalyst/H₂. Raney nickel may be recognized asa finely divided alloy of about 90% nickel and about 10% aluminum.“Raney” is a registered trademark of W. R. Grace and Company. Whenreduction is conducted with dihydrogen and using a catalyst, hydrogenmay be applied under a pressure from between about 1.4 bar and about 5bar, preferably under a pressure from between about 4 bar to about 5bar. A reducing agent may be used in an amount between about 1.2equivalents of reducing agent for every equivalent of protecteddinitrile to an excess of reducing agent, or between about 1.2equivalents of reducing agent for every equivalent of protecteddinitrile to about 8 equivalents of reducing agent for every equivalentof protected dinitrile.

The reaction may be conducted at a temperature at which the solvent usedis liquid. For example, if the solvent used is tetrahydrofuran, thereaction is conducted in a temperature range between about −108° C. toabout 67° C. In a preferred embodiment of the invention the reaction iscarried out in THF at a temperature of about 65° C.

Solvents are chosen based on the reducing agent used, and include, forexample, alcohols such as methanol and ethanol, acetic anhydride,dimethylformamide (DMF), tetrahydrofuran, diglyme, dimethoxyethane,toluene, and a mixture of alcohol and water. Alcohol solvents andmixtures of alcohol and water may optionally include liquid or gaseousammonia.

In one embodiment, a dinitrile protected by Boc groups is placed in anaqueous solution of ethanol and ammonia and hydrogenated in the presenceof Raney-nickel under a hydrogen atmosphere of about 4 bar to about 5bar. The reduction is allowed to proceed for about 15 hours at betweenabout 20° C. and about 25° C. Concentration to dryness affords aprotected diamine of Formula (II), where R is Boc, in greater than 95%yield.

In a further embodiment, a protected dinitrile as shown in Formula (IV)where R₁ is hydrogen and R₂ is phenyl, is added to a solution of about2.2 equivalents of lithium aluminum hydride in THF. By way of example,the reaction begins at about −30° C. and the reaction temperature israised over the course of about 80 minutes to about 20° C. A solution ofsodium hydroxide of about 4% concentration is added to the reactionuntil lithium aluminum hydride precipitates. A protected diamine ofFormula (I), where one of R₁ or R₂ is hydrogen and one of R₁ or R₂ isphenyl, remains in THF.

In yet another embodiment, for example, the reaction mixture containinglithium aluminum hydride in THF is treated with about one equivalent ofan alcohol. The alcohol may be, for example, ethanol or methanol and ispreferably methanol. The addition may take place at about 40° C. Wherethe alcohol used is methanol, the LiAlH₄ is converted to LiAlH₃OMe. Tothe LiAlH₃OMe is added a protected dinitrile as shown in Formula (IV),where one of R₁ or R₂ is hydrogen and one of R₁ or R₂ is phenyl, in THFat a temperature of about 40° C. followed by an aqueous quench,resulting in an increased yield of protected diamine and an eventualyield of greater than about 66% of tribenzaldehyde protectedintermediate. Purity is greater than about 99 area % (ionchromatography) when the protected diamine is treated with benzaldehydeas described below in Section D(2) and the resulting tribenzaldehydeprotected intermediate is crystallized and isolated. In anotherembodiment of the invention, the alcohol is in excess of the reducingagent.

An LAH reduction leading to the production of, for example can beconsidered as a four step process: the reduction itself, the aqueousquench and the removal of Al and Li salts, formation of the protectedtrientine, and crystallization. One such scheme for preparation ofbenylidene-(2-{3-[2-(benyzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl})-ethyl)-amineis set forth below.

In a certain preferred embodiment, the reducing agent LAH is treatedwith 1 equivalent of MeOH to increase yield and/or to facilitatefiltration. In yet another embodiment, the alcohol is in excess of thereducing agent. In a certain embodiment, the equivalents of the reducingagent and the alcohol are selected in a manner sufficient to ensurecomplete dissolution of the reaction product and/or to increase productyield or recovery. In a certain preferred embodiment, at least about 2.5equivalents of LAH and at least about 3.9 equivalents of MeOH are used.In another preferred embodiment, about 2.4 equivalents of LAH and atabout 4.0 equivalents of MeOH are used. In another preferred embodiment,about 2.5 equivalents of LAH and at about 3.95 equivalents of MeOH areused. Use of more than about 2.5 equivalents of LAH and more than about3.9 equivalents of MeOH, with the use of 2.7 equivalents of LAH and 3.95equivalents of MeOH being most preferred. Quench conditions usingstoichiometric amounts of MeOH are superior to aqueous conditions, whichcan lead to suspensions or viscous gels. In a preferred embodiment,coagulants NaOH and solid Na₂SO₄ are added to the reaction mixturefollowed by the addition of a saturated Na₂CO₃-solution to facilitatefiltration and recovery.

For crystallization and solvent exchange, in a certain embodiment, about2.2 equivalents of benzaldehyde are added to the reaction to obtain thetri-benzylidene-protected trientine, which is then crystallized fromisopropanol. Hexane may be used as an intermediate solvent to facilitatethe solvent exchange step in the reaction. In one preferred embodiment,crystallization of the protected intermediate (for example,tri-benzylidene-protected trientine) is carried out at a concentrationof approximately 4-5 L/mol to increase product yield.

D. Formation of a Triethylenetetramine Salt 1. Deprotection of ProtectedDiamine and Formation of Triethylenetetramine Salt

In another embodiment, amine protecting groups on a protected diamineare cleaved from a diamine by reaction of an acid with a protecteddiamine of Formula (II) or Formula (I), forming a triethylenetetraminesalt.

A triethylenetetramine salt with a triethylenetetramine to salt ratio ofabout 1 mole of trientine to about 3 moles of salt is formed by reactingbetween about 2 equivalents of acid to about 3 equivalents of acid withabout one equivalent of protected diamine. In one embodiment of theinvention, about 2.25 equivalents of hydrochloric acid are reacted withabout one equivalent of a protected diamine of Formula (I) to formtriethylenetetramine trihydrochloride salt. In another embodiment of theinvention, about 4 equivalents of succinic acid are reacted with aboutone equivalent of a protected diamine of Formula (II) to formtriethylenetetramine disuccinate salt.

A salt with a triethylenetetramine to salt ratio of about 1 mole oftriethylenetetramine to about 4 moles of salt or of about 1 mole oftriethylenetetramine to about 2 moles of salt is formed by reacting morethan about 3 equivalents of acid for each equivalent of protecteddiamine. The resulting ratio of moles of triethylenetetramine to salt isdetermined by the acid used. For example, hydrochloric acid formstriethylenetetramine tetrahydrochloride, while succinic acid formstriethylenetetramine disuccinic acid.

The hydrolysis reaction is conducted in a solvent. Solvents suitable forthe reaction include but are not limited to water, isopropanol, ethanol,methanol, or combinations thereof. Acids used in the reaction includebut are not limited to hydrochloric acid, hydrobromic acid, sulfuricacid, nitric acid, or phosphoric acid and the like, or organic acids,including but not limited to acetic acid, propanoic acid, hydroxyaceticacid, pyruvic acid, oxalic acid, malonic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,p-sulfonic acid, cyclamic acid, tartaric acid, succinic acid, malicacid, lactic acid, citric acid, maleic acid, salicyclic acid,p-aminosalicyclic acid, pamoic acid, or fumaric acid and the like. Othersuitable acids for the preparation of desired salts are described, forexample, in Handbook of Pharmaceutical Salts: Properties, Selection, andUse, P. Heinrich Stahl and Camille G. Wermuth (Eds.) (John Wiley & Sons,2002). Preferred acids for use in the reaction include fumaric acid,succinic acid, and maleic acid, which form triethylenetetraminefumarate, triethylenetetramine succinate, and triethylenetetraminemaleate salts. Succinic acid salts are preferred, and thetriethylenetetramine disuccinate salt anhydrate is most preferred. Ifhydrobromic acid or an organic acid is used in the reaction, heat isgenerally applied for salt formation to occur, although heat is notnecessary.

In one embodiment, a protected diamine of Formula (II), where R is Boc,may be prepared by reacting with an aqueous solution of isopropanol andabout 32% hydrochloric acid. Reaction at about 70° C. for about 30minutes cleaves the protecting groups, and triethylenetetramine isisolated by filtration in greater than 90% yield.

In another embodiment, a protected diamine of Formula (I), where one ofR₁ or R₂ is hydrogen and one of R₁ or R₂ is phenyl, is reacted with anexcess of aqueous hydrochloric acid to form triethylenetetraminetetrahydrochloride salt.

2. Protection of a Diamine, Purification of a Protected Diamine, andFormation of a Triethylenetetramine Salt

A protected diamine of Formula (I), in which, for example, R₁ may behydrogen and R₂ may be phenyl, is reacted with at least two equivalents,and preferably 2.2 equivalents, of an aldehyde or ketone including forexample but not limited to, formaldehyde, a substituted aromaticaldehyde, a substituted aliphatic aldehyde, a substituted aromaticketone, a substituted aliphatic ketone, or a substituted alkyl-aromaticketone to form the protected imidazolidine derivative of Formula (III),where R₁, R₂, R₃, R₄, R₅, and R₆ may be the same or different, and maybe hydrogen, an alkyl group including from one to twelve carbon atoms,aryl group, or an aromatic alkyl group. Optionally, additional additivesare also used, including but not limited to, for example, radicalscavengers. Radical scavengers include, for example, BHT(2,6-di-tert-butyl-4-methyl-phenol) and lithium chloride.

The reaction is conducted in a solvent. Solvents suitable for thereaction include but are not limited to water, isopropanol, ethanol,acetonitrile, methanol, or combinations thereof. In one embodiment,isopropanol is used as a solvent, resulting in a protected product thatprecipitates and is easily purified. The crystallized product isoptionally washed with a solvent, preferably isopropanol, andrecrystallized.

After the protected product is crystallized and purified the protectinggroups are cleaved by hydrolysis in the presence of an acid, forming atriethylenetetramine salt. As discussed above, the molar ratio oftriethylenetetramine to salt varies with the type and amount of acidused. Succinate salts are preferred, and most preferred is thedisuccinate salt.

In one embodiment, a solution in THF of protected diamine of Formula(I), where one of R₁ or R₂ is hydrogen and one of R₁ or R₂ is phenyl, istransferred by solvent exchange to isopropanol. About 2.2 equivalents ofbenzaldehyde are added to the reaction mixture over about two hours, andthe reaction is stirred for about 24 hours. After crystallization fromisopropanol, a product of the Formula (III) is formed, wherein R₁, R₃,and R₅ are hydrogen and R₂, R₄, and R₅ are phenyl. The crystallineproduct, a tribenzaldehyde protected intermediate, is isolated ingreater than 30% yield and at greater than 95% purity. Where, asdiscussed above, a protected diamine is reduced with lithium aluminumhydride that has been treated with an alcohol, yield of tribenzaldehydeprotected intermediate is greater than about 66%, with purity of greaterthan about 99% by gas chromatography.

Also provided are the formation of triethylenetetramine quaternarysalts. In an additional embodiment, a tribenzaldehyde protectedintermediate is reacted with at least more than about three equivalentsof aqueous hydrochloric acid, for example, to form triethylenetetraminetetrahydrochloride, which is precipitated in greater than about 90%yield and greater than about 98% purity from isopropanol.

In an additional embodiment, for example, a tribenzaldehyde intermediateis reacted with at least more than about three equivalents of aqueousmaleic acid to form triethylenetetramine tetramaleate, which isprecipitated from isopropanol.

In an additional embodiment, for example, a tribenzaldehyde intermediateis reacted with about three equivalents of aqueous hydrochloric acid toform triethylenetetramine trihydrochloride.

In an additional embodiment, for example, a tribenzaldehyde intermediateis reacted with about three equivalents of aqueous succinic acid to formtriethylenetetramine disuccinate.

In an additional embodiment, for example, a tribenzaldehyde intermediateis reacted with about three equivalents of aqueous maleic acid to formtriethylenetetramine trimaleate.

In an additional embodiment, for example, a tribenzaldehyde intermediateis reacted with about three equivalents of aqueous fumaric acid to formtriethylenetetramine trifumarate.

Primary salts of triethylenetetramine are also contemplated by theprocesses described herein.

In a certain embodiment, the reaction is facilitated by a reduction inreaction volume to 17 L/kg of product without the TBME extraction stepto ensure a high product yield. The reaction produced a fine-powderproduct.

In another embodiment, the reaction is facilitated by the addition ofsuccinic acid in methanol to a solution of tri-benzylidene-protectedtrientine in isopropanol/water. The resultant product is a sand-likeproduct following removal of methanol by distillation, cooling to about0° C., and subsequent recrystallization.

In a certain preferred embodiment, the removal of residual benzaldehydeis facilitated by dissolution of the product in H₂O at about 30° C.,followed by precipitation from H₂O/MeOH (¾) at about 0° C.

In a certain preferred embodiment, the removal of residual benzaldehydeis facilitated by dissolution of the product in H₂O at about 40° C.,followed by precipitation from H₂O/MeOH (4/1) at about 0° C.

In a certain preferred embodiment, the removal of residual benzaldehydeis facilitated by dissolution of the product in H₂O at about 30° C.,followed by precipitation from H₂O/iPrOH/MeOH (3/4/4) at about 0° C.

In a certain preferred embodiment, the removal of residual benzaldehydeis facilitated by dapting the volumes of the re-crystallizationprocedure to increase recovery.

In a certain embodiment, a reprecipitation is performed by the additionof alcohol.

3. Formation of a Triethylenetetramine Secondary Salt

In an additional embodiment, for example, a triethylenetetraminehydrochloride salt with molar ratio of about 1 mole triethylenetetramineto about two moles salt (that is, a secondary salt) is formed byreacting a triethylenetetramine hydrochloride salt with a molar ratio ofabout 1 mole triethylenetetramine to about 4 moles salt with a base in asolvent to produce free triethylenetetramine and free salt that isremoved as a precipitate. Optionally the precipitate is washed with asolvent, for example tert-butylmethylether. Free triethylenetetramine isthen reacted with at least about two equivalents of a concentrated acid,that is, an acid with a pH of less than about 1, to form atriethylenetetramine salt with a molar ratio of 1 mole oftriethylenetetramine to 2 moles of salt. The triethylenetetraminesecondary salt is precipitated from solution by addition of an alcohol.The triethylenetetramine secondary salt may be precipitated in highpurity without successive fractionation.

In a further embodiment, for example, the acid has been pre-dissolved ina solvent prior to its addition to the free triethylenetetramine.

Also provided herein is the synthesis of polymorphs oftriethylenetetramine salts.

Triethylenetetramine salts suitable for use in this embodiment includesalts of a molar ratio of triethylenetetramine to salt of about 1 toabout 4, including salts of the acids described above. A suitable saltis triethylenetetramine tetrahydrochloride. A particularly suitable saltis triethylenetetramine disuccinate, which is preferred.

Suitable bases for use in this embodiment include, for example, sodiummethoxide and sodium ethoxide. Suitable solvents include, for example,ethanol or methanol. Suitable acids include concentrated forms ofhydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, orphosphoric acid and the like, or organic acids, including, for example,acetic acid, propanoic acid, hydroxyacetic acid, pyruvic acid, oxalic(i.e., ethanedioic) acid, malonic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,cyclamic acid, tartaric acid, succinic acid, malic acid, lactic acid,citric acid, maleic acid, salicyclic acid, p-aminosalicyclic acid,pamoic acid, or fumaric acid and the like. Suitable alcohols forprecipitation of the product include, for example, ethanol, methanol,and isopropanol.

In one embodiment about 4 equivalents of sodium methoxide are mixed withabout 1 equivalent of triethylenetetramine tetrahydrochloride in amixture of methanol and ethanol, for example. The reaction mixture isfiltered, the solvent is evaporated, and the product is dissolved intert-butylmethylether and again filtered and dissolved, givingtriethylenetetramine in yield greater than about 95%. Thetriethylenetetramine is dissolved in about 2.0 equivalents ofconcentrated hydrochloric acid. Less than about 2 equivalents ofconcentrated hydrochloric acid may also be used. Ethanol is added andtriethylenetetramine dihydrochloride precipitates in yield greater thanabout 86%. Purity of triethylenetetramine dihydrochloride so produced isabout 100%, as determined using the methods set forth in USP27-NF22(page 1890) for analysis of trientine, with less than about 10 ppm heavymetals as determined by USP <231> II.

E. Purification of Triethylenetetramine

The compound of Formula (III) may be used for the preparation ofsubstantially pure triethylenetetramine. For example,triethylenetetramine in solution is reacted with between about 2equivalents and about 4 equivalents of a suitable aldehyde or ketone,including for example, formaldehyde, a substituted aromatic aldehyde, asubstituted aliphatic aldehyde, a substituted aromatic ketone, asubstituted aliphatic ketone, or a substituted alkyl-aromatic ketone toform the protected imidazolidine derivative of Formula (III), where R₁and R₂ may be the same or different, and may be for example but are notlimited to, hydrogen, an alkyl group including between one and twelvecarbon atoms, aryl group, or an aromatic alkyl group. In one embodiment,the aldehyde is benzaldehyde.

Suitable solvents for the reaction include but are not limited to water,isopropanol, ethanol, acetonitrile, methanol, or combinations thereof.In one embodiment, isopropanol is used as a solvent and the protectedproduct precipitates and is easily purified. The crystallized product isoptionally washed with a solvent, preferably isopropanol, andrecrystallized, further purifying the product. The protected product isdeprotected by reaction with an aqueous acid to form atriethylenetetramine salt as set forth in Section D(2), supra.

In another embodiment, triethylenetetramine is reacted with about 3.3equivalents of benzaldehyde in acetonitrile at 20° C. to form aprotected tribenzaldehyde intermediate. The protected tribenzaldehydeintermediate is crystallized from isopropanol and reacted in aqueoushydrochloric acid to form triethylenetetramine tetrahydrochloride inyield of about 90%.

Crystal production and analysis was performed on triethylenetetraminetetramaleate, triethylenetetramine tetrafumarate andtriethylenetetramine disuccinate synthesised according to syntheticschemes described herein. Crystals of x-ray quality fortriethylenetetramine tetramaleate may be grown, for example, by slowevaporation of a supersaturated solution of triethylenetetraminetetramaleate in water. The triethylenetetramine disuccinate andtriethylenetetramine tetrafumarate may be grown, for example, by slowevaporation of a solution of 12.58 mg triethylenetetramine disuccinateand 7.42 mg triethylenetetramine tetrafumarate in a water/ethanolmixture (1:1, 2 ml) over a period of 3 weeks. The structural coordinatesof Tables 1-3, and characterization of the crystalline structure, weredetermined based on measurements by x-ray powder diffraction.

The triethylenetetramine salts provided herein are of high purity.Triethylenetetramine salts may be produced with purity (calculated on adry basis) of, for example, at least about 80% triethylenetetraminesalt, at least about 85% triethylenetetramine salt, at least about 90%triethylenetetramine salt, at least about 95% triethylenetetramine salt,at least about 96% triethylenetetramine salt, at least about 97%triethylenetetramine salt, at least about 98% triethylenetetramine salt,at least about 99% triethylenetetramine salt, and about 100%triethylenetetramine salt. For example, triethylenetetramine succinatesalts, such as triethylenetetramine disuccinate, may also be producedwith purity (calculated on a dry basis) of, for example, at least about80% triethylenetetramine succinate salt, at least about 85%triethylenetetramine succinate salt, at least about 90%triethylenetetramine succinate salt, at least about 95%triethylenetetramine succinate salt, at least about 96%triethylenetetramine succinate salt, at least about 97%triethylenetetramine succinate salt, at least about 98%triethylenetetramine succinate salt, at least about 99%triethylenetetramine succinate salt, and about 100% triethylenetetraminesuccinate salt.

In addition to the compounds and salt forms provided herein, theinvention includes pharmaceutical compositions, including tablets,capsules, solutions, and suspensions for parenteral and oral deliveryforms and formulations, comprising a pharmaceutically acceptable carrierand therapeutically effective amounts of one or more of thetriethylenetetramine compounds herein provided. Pharmaceuticalcompositions including the triethylenetetramine disuccinate salt arepreferred, and pharmaceutical compositions includingtriethylenetetramine disuccinate anhydrate are most preferred.

In human and animal therapy for the treatment of undesired copperlevels, for example in the treatment of diabetes, cardiovasculardisease, and other disorders, diseases and conditions noted herein, thecompounds and their crystal forms described and provided herein, theirpharmaceutically acceptable salts, and pharmaceutically acceptablesolvates of either entity, can be administered alone, but will generallybe administered in admixture with a pharmaceutical carrier selected withregard to the intended route of administration and standardpharmaceutical practice. Preferably, they are administered orally in theform of tablets containing pharmaceutically acceptable excipients, suchas starch or lactose, or in capsules or ovules either alone or inadmixture with excipients, or in the form of elixirs, solutions orsuspensions containing flavouring or colouring agents. They can also beinjected parenterally, for example, intravenously, intramuscularly orsubcutaneously. For parenteral administration, they are best used in theform of a sterile aqueous solution which may contain other substances,for example enough salts or monosaccharides to make the solutionisotonic with blood. For buccal or sublingual administration they may beadministered in the form of tablets or lozenges which can be formulatedin a conventional manner.

Doses include those previously described. See Cooper, G. J., et al.,“Preventing and/or treating cardiovascular disease and/or associatedheart failure,” U.S. Pat. App. No. 2003/0203973, published Oct. 30,2003; and Cooper, G. J., et al., “Dosage forms and related therapies,”PCT Publication No. WO2004/017956, published Mar. 4, 2004. See also U.S.Pat. No. 6,897,243, which relates in part to the use oftriethylenetetramine in the treatment of diabetes.

For oral, parenteral, buccal and sublingual administration to patients,for example, the daily dosage level of the compounds herein and theirpharmaceutically acceptable salts and solvates may be from about 1 mg toabout 2400 mg per day (in single or divided doses). Other doses includedoses from about 5 mg to about 10 mg per day, from about 10 to about 50mg per day, from about 50 to about 100 mg per day, from about 100 mg toabout 200 mg per day, about from 200 mg to about 400 mg per day, fromabout 400 mg to about 600 mg per day, from about 600 mg to about 1200 mgper day, from about 600 mg to about 800 mg per day, from about 800 mg toabout 1000 mg per day, from about 1000 mg to about 1200 mg per day, andfrom about 1200 to about 2400 mg per day.

Thus, for example, tablets or capsules may contain from about 5 to about100 mg, and up to about 300 mg or more, of active compound foradministration singly, or two or more at a time, as appropriate. Thephysician will determine the actual dosage which will be most suitablefor an individual patient and it will vary with the age, weight andresponse of the particular patient. The above dosages are exemplary ofthe average case; there can, of course, be individual instances wherehigher or lower dosage ranges are merited and such are within the scopeof this invention.

Generally, in humans, oral administration of the compounds of theinvention is the preferred route. A preferred oral dosing regimen indiabetes and heart disease for a typical man is from about 400 mg toabout 1200 mg per day of compound when required. Preventative doses arelower, typically from about 1/10 to about 1/20 of the above amounts,including from about 20-40 mg to about 60-120 mg per day.

For veterinary use, a compound provided herein or a veterinarilyacceptable salt thereof, or a veterinarily acceptable solvate of eitherentity, is administered as a suitably acceptable formulation.

Thus the invention provides a pharmaceutical composition comprising atriethylenetetramine compound provided herein, or a pharmaceuticallyacceptable salt thereof, or a pharmaceutically acceptable solvate ofeither entity, together with a pharmaceutically acceptable diluent orcarrier.

It further provides a veterinary formulation comprising atriethylenetetramine compound provided herein, or a veterinarilyacceptable salt thereof, or a veterinarily acceptable solvate of eitherentity, together with a veterinarily acceptable diluent or carrier.

The invention also provides a triethylenetetramine compound providedherein, or a pharmaceutically acceptable salt thereof, or apharmaceutically acceptable solvate of either entity, or apharmaceutical composition containing any of the foregoing, for use as ahuman medicament.

In addition, it provides a triethylenetetramine compound providedherein, or a veterinarily acceptable salt thereof, or a veterinarilyacceptable solvate of either entity, or a veterinary formulationcontaining any of the foregoing, for use as an animal medicament.

In yet another aspect, the invention provides the use of atriethylenetetramine compound provided herein, or a pharmaceuticallyacceptable salt thereof, or a pharmaceutically acceptable solvate ofeither entity, for the manufacture of a human medicament for thecurative or prophylactic treatment of a medical condition for which acopper antagonist, particularly a copper (II) antagonist, is indicated.

It also provides the use of a triethylenetetramine compound providedherein, or a veterinarily acceptable salt thereof, or a veterinarilyacceptable solvate of either entity, for the manufacture of an animalmedicament for the curative or prophylactic treatment of a medicalcondition for which a a copper antagonist, particularly a copper (II)antagonist, is indicated.

Moreover, the invention includes use of the compounds and compositionsprovided herein for methods for treating and/or preventing, in whole orin part, various diseases, disorders and conditions, including but notlimited to glucose metabolism disorders; cardiovascular disorders;neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson'sdisease, and Huntington's disease); insulin disorders; liver disorders;lipid/cholesterol disorders; diseases, disorders, and conditions treatedor treatable with insulin; diseases, disorders, and conditions treatedor treatable with hypoglycemic agents; diseases, disorders, andconditions treated or treatable with statins and the like; diseases,disorders, and conditions treated or treatable with antihypertensiveagents; diseases, disorders, and conditions treated or treatable withanti-obesity agents; diseases, disorders or conditions treated ortreatable with biologically active protein C or a protein C derivative;and diseases, disorders, and conditions treated or treatable with copperantagonists including, for example, copper (II) chelators.

Diseases, disorders and conditions that may be treated include, forexample, atherosclerosis; peripheral vascular disease; cardiovasculardisease; heart disease; coronary heart disease; restenosis; angina;ischemia; heart failure; stroke; impaired glucose tolerance; impairedfasting glucose; prediabetes; diabetes and/or its complications,including type 1 and type 2 diabetes and their complications; insulinresistance; glucose metabolism diseases and disorders; chronichepatitis; fatty liver disease, including non-alcoholic and alcoholicfatty liver disease; steatohepatitis, including non-alcohlic andalcoholic steatohepatitis, and other conditions involving inflammationof the liver; Syndrome X; obesity and other weight related disorders;cardiomyopathy, including diabetic cardiomyopathy; hyperglycemia;hypercholesterolemia (e.g., elevated cholesterol in low-densitylipoprotein (LDL-C)); pre-hypertension, hypertension, secondaryhypertension, malignant hypertension, isolated systolic hypertension,and portal hypertension; hyperinsulinemia; hyperlipidemia; Alzheimer'sdisease Huntington's disease, and Parkinson's disease; degenerativediseases, including lupus and arthritis; nerve disease, includingdiabetic neuropathy; kidney disease, including diabetic nephropathy; eyedisease, including diabetic retinopathy and cataracts; acute coronarysyndromes, including myocardial infarction; vascular occlusivedisorders; diseases, disorders associated with a hypercoagulable stateor protein C deficiency, including but not limited to arterialthrombosis, arterial embolism, pulmonary embolism, deep venousthrombosis, venous thrombosis, renal vein thrombosis, mesenteric veinthrombosis, atheroembolic renal disease, thrombophlebitis, stroke, heartattack or angina, viral hemorrhagic fever, disseminated intravascularcoagulation, purpura fulminans, bone marrow and other transplantations,severe burns, major surgery, severe trauma, adult respiratory distresssyndrome, postphlebic syndrome, coumarin-induced skin necrosis;thrombotic diseases, disorders or conditions; sepsis and relateddiseases, disorders or conditions; diseases, disorders or conditionsrelating to undesired inflammation; thrombotic or embolic complicationsrelated to diseases, disorders or conditions including, but not limitedto, diabetes, hypertension, pre-hypertension, portal hypertension,hyperlipidemia, hypercholesteremia, and/or atherosclerosis.

Diseases, disorders and conditions that may be treated also include, forexample, (1) diseases, disorders and conditions characterized in part byany one or more of hyperlipidemia, hypercholesterolemia, hyperglycemia,hypertension, and/or hyperinsulinemia; (2) diseases, disorders orconditions characterized in whole or in part by (a) hypercupremia and/orcopper-related tissue damage and (b) hyperglycemia, insulin resistance,impaired glucose tolerance, and/or impaired fasting glucose, and/orelevated or undesired levels of LDL-C, or predisposition to, or riskfor, (a) and (b); (3) diseases, disorders and conditions characterizedin whole or in part by (a) excess copper and/or copper-related tissuedamage and (b) a BMI from about 25 to about 29.9 or a BMI greater thanabout 30 (including subjects having a BMI from about 30 to about 34.9(obesity class I), from about 35 to 39.9 (obesity class II), and greaterthan about 40 (obesity class III)); (4) diseases, disorders orconditions characterized in whole or in part by (a) excess copper and/orcopper-related tissue damage, and (b) protein C deficiency and/orundesired coagulation activity, or predisposition to, or risk for, (a)and (b); (5) diseases, disorders or conditions characterized in whole orin part by (a) excess copper and/or copper-related tissue damage, and(b) excess body fat; and subjects within the World Health Organization(WHO) classification for overweight and obesity (includingsubclassifications based on race and waist circumference), who are atrisk for comorbid conditions, including hypertension, type 2 diabetesmellitus, and cardiovascular disease.

The invention includes methods for treating a subject having orsuspected of having or predisposed to, or at risk for, for example, anydiseases, disorders and/or conditions described or referenced herein.Such compounds may be administered in amounts, for example, that areeffective to (1) decrease body and/or tissue copper levels, (2) increasecopper output in the urine of said subject, and/or (3) decrease copperuptake, for example, in the gastrointestinal tract. Such compositionsinclude, for example, tablets, capsules, solutions and suspensions forparenteral and oral delivery forms and formulations.

The invention includes methods for administering a therapeuticallyeffective amount of a triethylenetetramine compound provided herein in adelayed release preparation, a slow release preparation, an extendedrelease preparation, a controlled release preparation, and/or in arepeat action preparation. Such preparations may be administered to asubject having or suspected of having or predisposed to diseases,disorders and/or conditions referenced herein. Such compounds may beadministered in amounts, for example, that are effective to (1) decreasebody and/or tissue copper levels, (2) increase copper output in theurine of said subject, (3) decrease copper uptake, for example, in thegastrointestinal tract, and/or (4) lower LDL-C. Such compositionsinclude, for example, tablets, capsules, solutions and suspensions forparenteral and oral delivery forms and formulations.

The invention also includes pharmaceutical compositions, includingtablets and capsules and other oral delivery forms and formulations,comprising a pharmaceutically acceptable carrier and therapeuticallyeffective amounts of a 3-hydroxy-3-methylglutaryl coenzyme A reductaseinhibitor and a triethylenetetramine compound as provided herein, withtriethylenetetramine disuccinate anhydrate being preferred, particularlyin the disclosed crystal form. Suitable 3-hydroxy-3-methylglutarylcoenzyme A reductase inhibitors include the statins. Preferred statinsare simvastatin, atorvastatin, lovastatin, pravastatin, fluvastatin, androsuvastatin. Other statins include itavastatin and visastatin.3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors are presentin the compositions of the invention amounts, for example, that areeffective to lower LDL-C. The pharmaceutical compositions may beadministered in amounts, for example, that are effective to (1) decreasebody and/or tissue copper levels, (2) increase copper output in theurine of said subject, (3) decrease copper uptake, for example, in thegastrointestinal tract, and/or (4) lower LDL-C. The pharmaceuticalcompositions may be used for treating and/or preventing, in whole or inpart, various diseases, disorders and conditions, including, forexample, atherosclerosis; coronary heart disease; impaired glucosetolerance; impaired fasting glucose; diabetes and/or its complications,including type 1 and type 2 diabetes and their complications; insulinresistance; Syndrome X; obesity and other weight related disorders;cardiomyopathy, including diabetic cardiomyopathy; hyperglycemia,hypercholesterolemia (e.g., elevated cholesterol in low-densitylipoprotein (LDL-C)), hypertension, hyperinsulinemia, and/orhyperlipidemia; diseases, disorders and conditions characterized in partby any one or more of hyperlipidemia, hypercholesterolemia,hyperglycemia, hypertension, and/or hyperinsulinemia; and, diseases,disorders or conditions characterized in whole or in part by (a)hypercupremia and/or copper-related tissue damage and (b) hyperglycemia,insulin resistance, impaired glucose tolerance, and/or impaired fastingglucose, and/or elevated or undesired levels of LDL-C, or predispositionto, or risk for, (a) and (b).

The invention also includes pharmaceutical compositions, includingtablets and capsules and other oral delivery forms and formulations,comprising a pharmaceutically acceptable carrier and therapeuticallyeffective amounts of a hypoglycemic agent and a triethylenetetraminecompound as provided herein, with triethylenetetramine disuccinateanhydrate being preferred, particularly in the disclosed crystal form.Suitable hypoglycemic agents include biguanides (for example,metformin), thiazolidinediones (for example, troglitazone,rosiglitazone, and pioglitazone), α-glucosidase inhibitors (for example,acarbose and miglitol), and sulfonylureas (for example, tolbutamide,chlorpropamide, gliclazide, glibenclamide, glipizide, and glimepiride).Other hypoglycemic agents include amylin and amylin agonists (e.g.,pramlintide, which is ^(25,28,29)Pro-h-amylin), GLP-1 and GLP-1 agonists(e.g., Arg(34)Lys(26)-(N-ε-(γ-Glu(N-α-hexadecanoyl))-GLP-1(7-37), orGLP-1LA)), and exendin and exendin agonists (e.g., exendin-4). Suchcompounds may be present in the compositions of the invention inamounts, for example, that are effective to (1) lower blood glucose, (2)lower serum glucose, (3) lower urine glucose, (4) lower glycosylatedhemoglobin (HbA_(1c)) levels, (5) lower fructosamine, (6) lowerpostprandial glycemia, (7) ameliorate impaired glucose tolerance, (8)ameliorate impaired fasting glucose, and/or (9) lower the rate and/orseverity of hypoglycemic events, including severe hypoglycemic events.The pharmaceutical compositions may be used for treating and/orpreventing, in whole or in part, various diseases, disorders andconditions, including, for example, impaired glucose tolerance; impairedfasting glucose; diabetes and/or its complications, including type 1 andtype 2 diabetes and their complications; insulin resistance; Syndrome X;obesity and other weight related disorders; cardiomyopathy, includingdiabetic cardiomyopathy; nerve diseases, including diabetic neuropathy;kidney disease, including diabetic nephropathy; eye disease, includingdiabetic retinopathy and cataracts; hyperglycemia, hypercholesterolemia,hypertension, hyperinsulinemia, hyperlipidemia, atherosclerosis, and/ortissue ischemia, and diseases and disorders characterized at least inpart by any one or more of hyperglycemia, hypercholesterolemia,hypertension, hyperinsulinemia, hyperlipidemia, atherosclerosis, andtissue ischemia; neurodegenerative disorders, including Alzheimer'sdisease and Parkinson's disease; and, diseases, disorders or conditionscharacterized in whole or in part by (a) hypercupremia and/orcopper-related tissue damage and (b) hyperglycemia, insulin resistance,impaired glucose tolerance, and/or impaired fasting glucose, orpredisposition to, or risk for, (a) and (b).

The invention also includes pharmaceutical compositions, includingtablets and capsules and other oral delivery forms and formulations,comprising a pharmaceutically acceptable carrier and therapeuticallyeffective amounts of an antihypertensive agent and atriethylenetetramine compound as provided herein, withtriethylenetetramine disuccinate anhydrate being preferred, particularlyin the disclosed crystal form. Suitable antihypertensive agents includeare those that lower blood pressure and include, for example, diuretics(including hydrochloride and chlorthalidone), α-adrenergic receptorantagonists (including prazosin, terazosin, doxazosin, ketanserin,indoramin, urapidil, clonideine, guanabenz, guanfacine, guanadrel,reserpine, and metyrosine), β₁-selective adrenergic antagonist(including metoprolol, atenolol, esmolol, acebutolol, bopindolol,carteolol, oxprenolol, penbutolol, medroxalol, bucindolol, levobunolol,metipranolol, bisoprolol, nebivolol, betaxolol, celiprolol, sotalol,propafenone, propranolol, timolol maleate, and nadolol), ACE inhibitors(including captopriol, fentiapril, pivalopril, zofenopril, alacepril,enalapril, enalaprilat, enalaprilo, lisinopril, benazepril, quinapril,moexipril), calcium channel blockers (including nisoldipine, verapamil,diltiazem, nifedipine, nimodipine, felodipine, nicardipine, isradipine,amlodipine, and bepridil), angiotensin II receptor antagonists(including losartan, candesartan, irbesartan, valsartan, telmisartan,eprosartan, and olmesartan medoxomil), and vasodilators (includinghydralazine, Minoxidil, sodium nitroprusside, diazoxide, bosentan,eporprostenol, treprostinil, and iloprost). Other antihypertensiveagents include sympatholytic agents (e.g., methyldopa), ganglionicblocking agents (including mecamylamine and trimethaphan), andendothelin receptor antagonists (including bosentan and sitaxsentan).The compounds may be present in amounts, for example, that are effectiveto (1) decrease body and/or tissue copper levels, (2) increase copperoutput in the urine of said subject, (3) decrease copper uptake, forexample, in the gastrointestinal tract, and/or (4) lower blood pressure.The invention also relates to methods of using such compositions totreat subjects suffering from or at risk for various diseases,disorders, and conditions, including pre-hypertension, hypertension(including essential hypertension and grades 1, 2 and 3 hypertension)and related cardiovascular diseases; secondary hypertension; malignanthypertension; isolated systolic hypertension; atherosclerosis; coronaryheart disease; impaired glucose tolerance; impaired fasting glucose;diabetes, including type 1 and type 2 diabetes, and their complications;insulin resistance; Syndrome X; obesity and other weight relateddisorders; cardiomyopathy, including diabetic cardiomyopathy; diseasesand disorders characterized in part by any one or more of hypertension,hyperlipidemia, hypercholesterolemia (e.g., elevated cholesterol inlow-density lipoprotein (LDL-C)), hyperglycemia, and/orhyperinsulinemia; and, characterized in whole or in part by (a)hypercupremia and/or copper-related tissue damage and (b) hypertension,insulin, or predisposition to, or risk for, (a) and (b). The inventionincludes methods for the use of therapeutically effective amounts of atriethylenetetramine compound provided herein in the manufacture of amedicament. Such medicaments include, for example, tablets, capsules,solutions and suspensions for parenteral and oral delivery forms andformulations. Such medicaments include those for the treatment of asubject as disclosed herein.

The compounds of the invention, particularly triethylenetetraminedisuccinate anhydrate, for example, in the disclosed crystal form, mayalso be prepared with an anti-obesity agent or an insulin.

Doses for such triethylenetetramine compounds, salts and/or solvates asprovided herein are envisage to be administered in a therapeuticallyeffective amount, for example, to lower copper values in a subject.

The compounds of the invention may also be pre-complexed with anon-copper metal ion prior to administration for therapy. Metal ionsused for pre-complexing are pharmaceutically acceptable and have a lowerassociation constant for the copper antagonist than that of copper. Forexample, a metal ion for pre-complexing a copper antagonist thatchelates Cu²⁺ is one that has a lower binding affinity for the copperantagonist than Cu²⁺. Preferably, the non-copper metal ion has anassociation constant for triethylenetetramine that is equal to or lessthan about 10⁻¹⁹, more preferably less than or equal to about 10⁻¹⁸,still more preferably less than or equal to about 10⁻¹⁵, even morepreferably less than or equal to about 10⁻¹², 10⁻¹⁰, or 10⁻⁹, and mostpreferably less than or equal to about 10⁻⁸, 10⁻⁷ or 10⁻⁵.

Preferred metal ions for precomplexing include, for example, calcium(e.g., Ca²⁺), magnesium (e.g., Mg²⁺), chromium (e.g., Cr²⁺ and Cr³⁺),manganese (e.g., Mn²⁺), zinc (e.g., Zn²⁺), and iron (e.g., Fe²⁺). Mostpreferred metal ions for precomplexing are calcium, zinc, and iron.Other metals include, for example, cobalt (e.g., Co²⁺), nickel (e.g.,Ni²⁺), silver (e.g., Ag¹⁺), and selenium (e.g., Se⁴⁺). Non-copper metalsare chosen with regard, for example, to their relative binding to thetriethylenetetramine, the dose of the triethylenetetramine to beadministered, and relative to potential toxicity following displacementof the non-copper metal ion. Examples of pre-complexedtriethylenetetramines include triethylenetetramine disuccinatepre-complexed with a metal ion having a binding constant lower thancopper, for example, triethylenetetramine disuccinate pre-complexed withzinc or calcium (e.g., Zn²⁺ and Ca²⁺). Without intending to be bound toany particular mechanism or mode of action, precomplexing allows lowerdosing.

The invention includes methods for the use of a therapeuticallyeffective amount of a triethylenetetramine compound provided herein inthe manufacture of a dosage form. Such dosage forms include, forexample, tablets, capsules, solutions and suspensions for parenteral andoral delivery forms and formulations. Such dosage forms include thosefor the treatment of a subject as disclosed herein.

The invention includes an article of manufacture comprising a vesselcontaining a therapeutically effective amount of a triethylenetetraminecompound provided herein and instructions for use, including use for thetreatment of a subject.

The invention includes an article of manufacture comprising packagingmaterial containing one or more dosage forms containing atriethylenetetramine compound provided herein, wherein the packagingmaterial has a label that indicates that the dosage form can be used fora subject having or suspected of having or predisposed to any of thediseases, disorders and/or conditions described or referenced herein.Such dosage forms include, for example, tablets, capsules, solutions andsuspensions for parenteral and oral delivery forms and formulations.

In yet another aspect of this invention is a kit comprising (a) at leastone triethylenetetramine compound, or salt or crystal thereof, and apharmaceutically acceptable carrier, excipient and/or additive in a unitdosage form, and (b) means for containing the unit form. Since thepresent invention has an aspect that relates to the treatment of thedisease/conditions described herein with a combination of activeingredients, the invention also relates to combining separatepharmaceutical compositions in kit form. A kit may contain apharmaceutical composition comprising triethylenetetramine compound, orsalt or crystal thereof, as provided herein, either alone or togetherwith a second compound as described herein.

The kit comprises means for containing the composition such as acontainer, a bottle or a foil packet. Typically the kit comprisesdirections for the administration of the composition.

An example of such a kit is a so-called blister pack. Blister packs arewell known in the packaging industry and are being widely used for thepackaging of pharmaceutical unit dosage forms (tablets, capsules, andthe like). Blister packs generally consist of a sheet of relativelystiff material covered with a foil of a preferably transparent plasticmaterial. During the packaging process recesses are formed in theplastic foil. The recesses have the size and shape of the tablets orcapsules to be packed. Next, the tablets or capsules are placed in therecesses and the sheet of relatively stiff material is sealed againstthe plastic foil at the face of the foil which is opposite from thedirection in which the recesses were formed. As a result, the tablets orcapsules are sealed in the recesses between the plastic foil and thesheet. Preferably the strength of the sheet is such that the tablets orcapsules can be removed from the blister pack by manually applyingpressure on the recesses whereby an opening is formed in the sheet atthe place of the recess. The tablet or capsule can then be removed viasaid opening. It may be desirable to provide a memory aid on the kit,e.g., in the form of numbers next to the tablets or capsules whereby thenumbers correspond with the days of the regimen which the tablets orcapsules so specified should be ingested. Another example of such amemory aid is a calendar printed on the card, e.g., as follows “FirstWeek, Monday, Tuesday, . . . etc. . . . Second Week, Monday, Tuesday, .. . ” etc. Other variations of memory aids will be readily apparent. A“daily dose” can be a single tablet or capsule or several pills orcapsules to be taken on a given day.

In another specific embodiment of the invention, a dispenser designed todispense the daily doses one at a time in the order of their intendeduse is provided. Preferably, the dispenser is equipped with amemory-aid, so as to further facilitate compliance with the regimen. Anexample of such a memory-aid is a mechanical counter which indicates thenumber of daily doses that has been dispensed. Another example of such amemory-aid is a battery-powered micro-chip memory coupled with a liquidcrystal readout, or audible reminder signal which, for example, readsout the date that the last daily dose has been taken and/or reminds onewhen the next dose is to be taken.

The invention includes a formulation comprising a triethylenetetraminecompound provided herein in amounts effective to remove copper from thebody of a subject and reduce elevated copper levels. Such formulationsinclude, for example, tablets, capsules, solutions and suspensions forparenteral and oral delivery forms and formulations.

The following experiments set forth in the following examples areillustrative of the present inventions and are not intended to limit theinventions.

EXAMPLES

Raw materials used in the Examples were purchased from Fluka Company(Switzerland) and Aldrich Chemicals (Milwaukee, Wis.). Product puritywas assessed by thin-layer chromatography and/or NMR. In some cases agas chromatography analysis was performed. Qualitative thin layerchromatography (TLC) was generally used for in process controls tomonitor conversions.

TLC parameters for those examples related to Boc were as follows:

TLC-plate: SiO₂ F60 Eluent: Methanol Detection 1) 0.2 g Ninhydrin in 100ml ethanol 2) Heating to 110° C. R_(f)-values: Ethylenediamine (1) R_(f)= 0.0-0.30 (yellow) Dinitrile (2) R_(f) = 0.45-0.48 (orange/pink)Boc-protected Dinitrile (3) R_(f) = 0.49-0.52 (light brown)Boc-protected Diamine (4) R_(f) = 0.19-0.21 (brown) TriethylenetetramineR_(f) = 0.0-0.10 (yellow)

Gas chromatography analysis was used to assess the purity of thebenzaldehyde derivatized intermediates. The gas chromatography analysiswas combined with NMR, and was not used to monitor conversions due tocolumn degradation after a few injections.

Gas chromatography parameters were as follows:

Column: DB-5, 15 m × 0.25 mm i.d. × 0.25 μm film Mobile Phase: H₂ @ 1.5mL/min const. flow Time Temp. Heating Rate Temp. program: [min] [° C.][° C./min] 0 40 0 1.2 40 0 4.2 70 10 13.2 300 33.3 17.2 300 0 InjectorTemp. 250° C. Detector Temp. 300° C. (FID) Detection: Step: AutoAverage: on Split flow/Split Ratio: 75 mL/min/1:50 Injection Volume: 1.0μL

Example 1

Example 1 describes the preparation of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile.

Ethylenediamine hydrochloride salt (66.5 g, 0.5 mol, 1.0 eq) wasdissolved in water (350 ml). KCN (65.1 g, 1.0 mol, 2.0 eq) was added tothe reaction mixture endothermically. A solution of formaldehyde (36.5%in water) (83 ml, 1.1 mol, 2.2 eq) was added to the reaction mixtureover about 75 min, so that the internal temperature stayed below about25° C. The reaction mixture was cooled with an ice bath. After theaddition of 50% of the formaldehyde solution the pH was brought fromabout 12 to about 10 by adding acetic acid.

After complete addition of the formaldehyde solution the pH was adjustedto about 5 with acetic acid. The reaction mixture was slowly stirred at20° C. for 17 hours. Thin layer chromatography showed completeconversion. Benzaldehyde (53.05 g, 50.8 mL, 0.5 mol, 1.0 eq) was added,and the reaction mixture was stirred at 20° C. for 40 min. The reactionmixture was extracted with ethyl acetate (100 mL). The aqueous phase wasextracted 3 times with ethyl acetate (3×100 mL).

The combined organic phases were concentrated to dryness on a rotaryevaporator to afford 203 g of crude(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile. The crudeproduct was crystallized from n-butanol/cyclohexane (125 mL/75 mL). Theproduct was filtered, washed with n-butanol/cyclohexane (35 mL/35 mL)and dried on a rotary evaporator (external temperature: 40° C., p=20mbar). 22.7 g (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrilewas obtained as a white solid, corresponding to a yield of 20%. Thepurity was 99.5 area % as measured by gas chromatography. The ¹H-NMRspectrum of (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile isshown in FIG. 4.

Example 2

This example demonstrates another preparation of a3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile.

1 equal molar of ethylenediamine and 2 equal moles of KCN were mixed ina reaction vessel pre-purged with nitrogen gas. Water was added to thereaction mixture and the feeding vessel was rinsed with water. Thefeeding vessel was then charged with 2 equal moles of HCl (32%) and thesolution was then slowly added to the mixture so that the internaltemperature of the reaction did not exceed 25° C. Following the HCladdition, the reaction mixture was cooled to about 3° C. 2 equal molesof formaldehyde solution (30% dissolved in water) was added to thereaction mixture with cooling over 40 minutes. The reaction mixture wasthen stirred for 2.5 hours at a temperature range of 15° C. to 20° C.For NMR analysis, 5-10 mg of the reaction mixture was dissolved in 1000μL D2O and analyzed by proton-NMR. The spectrum indicated completedisappearance of ethylenediamine. Thirty minutes after the addition offormaldehyde, a 0.5 equal molar of NaH₂PO₄ solution was added to thereaction mixture at an internal reaction temperature of between 10° C.to 12° C. The resultant pH of the reaction mixture dropped to about 6.N-butyl alcohol was added to the reaction mixture at an internaltemperature of 11.6° C. Thirty minutes after the addition of n-butylalcohol, 1 equal mole of benzaldehyde was added to the reaction mixtureat an internal reaction temperature of 11° C. to 12° C. After half ofthe benzaldehyde has been added,3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile seed crystalswere added to the solution. The reaction mixture was then cooled to −4°C. over the span of 4 hours then stirred at −4° C. for 11.5 hours. Thefilter cake was washed with deionized water at 15-19° C., then washedwith n-butyl alcohol at 0° C. in two portions; the reaction mixture wasthen washed with isopropyl alcohol at −2° C. Crude(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile was transferredinto a PROVATEC-dryer and dried at 50° C. under 4 mbar of pressure. Thereaction produced(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile at 99.31%purity, at 86.7% yield.

In another run, the reaction produced(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile at 99.63%purity, at 85.1% yield.

Example 3

This Example demonstrates the preparation ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine.

A solution of LiAlH₄ (34.5 mmol, 3.0 eq) in THF (112.5 mL) was cooled to−30° C. A solution of the compound of Example 1,(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (2.6 g, 11.5mmol, 1.0 eq), in THF (78 mL), was added over about 25 min. The reactionmixture was stirred at −30° C. for 10 min and then warmed to 20° C. over80 min. A while solid precipitated. Thin-layer chromatography showedcomplete consumption of the starting material. Diethyl ether (8 mL) wasadded to the reaction mixture. Water (6 mL) was exothermically added tothe reaction mixture at 0° C. over 20 min, and H₂ evolution occurred.Benzaldehyde (2.7 g, 2.6 mL, 25.3 mmol, 2.2 eq) was added at 20° C. andthe reaction mixture stirred for 45 min.

The suspension was filtered and washed twice with diethyl ether (2×15mL). The filtrate was concentrated to dryness to afford 4.0 g of crudebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine.The crude product was recrystallized from acetonitrile. Filtration at 0°C. and drying on a rotary evaporator gave 1.2 g ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas a white solid (purity: 95.8 area % by gas chromatography). A secondcrystallization from the mother liquor gave 0.3 g of product. The totalyield was 32%. The ¹H-NMR spectrum of thebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminein CDCl₃ is shown in FIG. 5.

Example 4

This example demonstrates another preparation of abenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine.

1 equal mole of (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrileand THF were mixed in a reactor and the reactor was purged with nitrogengas. The reaction mixture was then transferred to a feeding vessel. Thereactor was washed with THE and the solution was added into the mixturein the feeding vessel. 3 equal moles of LiAlH₄ in THF solution was addedto the reactor. 4 equal moles of methanol and THF were mixed in anotherfeeding vessel, this mixture was added over the next 40 minutes at aninternal temperature of 35 to 40° C. into the main reaction mixture. Thesolution was then heated to an internal temperature of 40° C. underconstant stirring for one additional hour. The mixture of methanol andTHF was slowly added to the mixture in the feeding vessel at an internaltemperature of 39° C.-45° C. The mixture was stirred at an internaltemperature of 40° C. for an additional 30 minutes. The mixture was thencooled to an internal temperature of 10° C. within 1 hour. Na₂SO₄ andNaOH were added to the reaction mixture. Saturated Na₂CO₃-solution wasadded at an internal temperature of 10-18° C. via a feeding vessel. Thereaction mixture was stirred at an internal temperature of 10° C.overnight. Na₂HPO₄ was added to the mixture and the reaction suspensionwas filtered into the stirring vessel. The filter cake was washed withtwo volumes of THF. The suspension of Na2HPO4 and the filtrate wasstirred for one hour in the stirring vessel and then filtered over anutsch. The filter cake was washed with THF. Both filtrates (filtrationand washing) were then transferred into the second reactor (viainline-filter). 2 equal moles of benzaldehyde was added to the filtrateat 20° C. (via inline filter). THF was distilled off at an internaltemperature of 10-20° C. and at a pressure of 130-210 mbar, externaltemperature (ET)=50° C. Hexane was then added (ET=40° C.) to themixture. Water was then distilled off at an internal temperature of16-20° C., with a pressure of 170-190 mbar and a AT of 40-43° C. Thehexane was further distilled off at an internal temperature of 16° C.,at a pressure of 160-180 mbar and ET=44° C. Isopropanol was added to thereactor via a feeding vessel. Isopropanol was distilled off at IT=10-27°C., p=30-114 mbar and ET=50° C. The reaction mixture was stirred for anhour at an internal temperature of 35° C. The reaction mixture was thencooled to −5° C. and stirred overnight. The nutsch was again purged withnitrogen and the suspension was again filtered. The filter cake waswashed with cold (6° C.) isopropanol. The product,benzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine,was allowed to dry in a PROVATECH dryer at a temperature of 30-40° C. ata pressure of less than or equal to 30 mbar. The reaction was carriedout in five independent reaction vessels with the following results:Flask 1 gavebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of 99.93% as measured by gaschromatography, which represented a total yield of 84.85%. Flask 2 gavebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of 99.42% as measured by gaschromatography, which represented a total yield of 77.1%. Flask 3 gavebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of 99.89% as measured by gaschromatography, which represented a total yield of 78.3%. Flask 4 gavebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of 99.87% as measured by gaschromatography, which represented a total yield of 82.7%.

Example 5

This Example demonstrates the preparation of a triethylenetetraminetetrahydrochloride.Benzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine(3.15 g, 7.67 mmol), was dissolved in water (5 mL) and concentrated HCl(5 ml) and stirred for 5 min at 20° C. The reaction mixture wasextracted once with TBME (10 mL). The aqueous phase was treated withisopropanol (30 mL), whereupon the product, triethylenetetraminetetrahydrochloride, precipitated. The product was filtered, washed withisopropanol (10 mL) and dried on a rotary evaporator. 2.01 g ofcrystallized triethylenetetramine tetrahydrochloride was obtained as awhite solid. FIG. 6 shows an ¹H-NMR spectrum of triethylenetetraminetetrahydrochloride in D₂O.

Example 6

This Example demonstrates a dinitrile reduction of an intermediate thatwas protected by, for example, a benzaldehyde protecting group. Abenzaldehyde protecting group was be used as shown in Scheme 1, in whicha solution of a [2-(cyanomethyl-amino)-ethylamino]-acetonitrileintermediate (2) was treated with 1 equivalent of benzaldehyde andstirred at 20° C. for about 2 hours. Work-up resulted in crude(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) in 84%yield. Purification by crystallization from cyclohexane resulted in(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) in 40%yield.

The synthesis of [2-(cyanomethyl-amino)-ethylamino]-acetonitrileintermediate (2) was accomplished by a Strecker-type synthesis as shown,for example, in Scheme 2.

A solution of 1 equivalent of ethylenediamine hydrochloride salt inwater was treated with 2 equivalents of KCN. A solution of 2.2equivalents of formaldehyde in water was added over about 75 minutes andthe pH was adjusted to 5 with acetic acid. Thin-layer chromatographyshowed complete conversion after about 17 hours at 20° C. Theintermediate (2) was not isolated but was treated with 1 equivalent ofbenzaldehyde. Extraction with ethyl acetate and crystallization frombutanol/cyclohexane resulted in(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) in 20% yieldwith a purity of 99.5 area % by gas chromatography. The crystallinity of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) allowedsimple purification of the intermediate.

Alternatively, a dinitrile intermediate is treated with sodium phosphatemonobasic (NaH₂PO₄) and benzaldehyde at a temperature of about 10° C.and a pH of about 6 to 7, followed by addition of n-butylalcohol,resulting in the production of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5).

Example 7

This Example demonstrates the reduction of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) with LiAlH₄and protection using benzaldehyde. As shown in Scheme 3, a solution of 1equivalent of (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile(5) was added to a solution of 3 equivalents of LiAlH₄ in THF at about−30° C. The reaction mixture was warmed to about 20° C. over 80 minutes,whereupon thin-layer chromatography showed complete conversion. Thereaction mixture was treated with water (vigorous, exothermic reactionwith H₂-evolution) and aluminum salts were removed by filtration.Isolation of the amine intermediate2-[3-(2-Amino-ethyl)-2-phenyl-imidazolidin-1-yl]-ethylamine (8) as apure compound was not readily achieved. To add protecting groups andallow isolation the amine intermediate (8) was reacted in situ with 2.2equivalents of benzaldehyde after aqueous quench. After work-up andcrystallization from acetonitrile, crystallinebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine(7) was obtained in 32% yield and with a purity of 95.8 area % by gaschromatography.

Example 8

This Example demonstrates the preparation of crystalline intermediatebenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine(7). This preparation is significant, for example, because thecrystalline intermediate (7) may be hydrolyzed in the presence of anacid to form a triethylenetetramine salt. Hydrochloric acid is used forthe production of triethylenetetramine tetrahydrochloride, as shown inScheme 4. The formation of triethylenetetramine dihydrochloride is shownin Scheme 5. The use of succinic acid for the production of adisuccinate salt is shown in Scheme 6. The use of fumaric acid andmaleic acid to produce triethylenetetramine tetrafumaric salt andtriethylenetetramine tetramaleic salt, respectively, is depicted inScheme 7 and Scheme 8, respectively.

Salts formed from succinic acid and maleic acid exhibitednon-hygroscopic properties. Phosphate, mesylate, benzoate, citrate, andmalate salts exhibited low crystal formation, sulfate salt exhibited alow melting point, and tartrate salt was hygroscopic. Salts in allmanner of stoichiometric and nonstoichiometric ratios are contemplatedby the invention, including salts present in an acid totriethylenetetramine ratio of 1 to 1, 2 to 1, 3 to 1, and 4 to 1. Theproperties of a number of triethylenetetramine salts produced by amethod of the invention are set forth in Table 1. All of the salts setforth in Table 1 were found to be colorless, and the salts of thehydrochloric, succinic, and maleic acid were obtained as free flowingcrystalline powders. The salt of fumaric acid was obtained as a woolysolid. The fumarate, succinate, and maleic acid salts were exposed to80° C. for nine days, and the IR, ¹H-NMR, and DSC after that time showedno changes.

TABLE 1 Stoichiometry n_(acid):n_(trientine) Triethylene- Bulk Meltingpoint Solubility Yield ¹H-NMR; EL. tetramine Density [° C.] DSC (water)Salt [%] Analysis [% w/w] [g/cm³] (onset/peak) [g/L] pH Fumaric 83 4:123.9 0.09 186.01/187.97 10.3 3.32 (Sat. Sol'n) Maleic 78 4:1 23.9 0.19180.34/181.86 10.8 2.91 (Sat. Sol'n) Succinic 87 2:1 38.2 0.33180.05/179.91 >389 5.22 (0.16M) Hydrochloric 82 2:1 66.7 0.21122.53/122.92 very 8.0 soluble (0.02M)

Salts of the invention were also analyzed for hygroscopicity, as shownin Table 2, after exposure to a controlled humidity atmosphere (38.6%relative humidity, 19° C.) for several days. The succinate and themaleic acid salts may be considered non-hygroscopic salts. Thedihydrochloride salt absorbed water in a reversible fashion; the DSC ofa dried sample was identical to that of a sample with almost 0% watercontent. The fumarate salt absorbed water to a constant amount of about3.51%, which was interpreted as the formation of a monohydrate. In someembodiments of the invention, seed crystals may be added to aid saltcrystal formation.

TABLE 2 Results of Karl Fischer Water Determination (in % assay) initial24 hours 3 days 7 days Dihydrochloride Sample 1 0.07 0.24 10.63  —Comparison 0.21 Maleate Sample 1 0.04 0.04 0.04 0.03 Comparison — — —Succinate Sample 1 0.01 0.02 0.02 0.01 Comparison — — — Fumarate Sample1 1.00 3.51 3.51 Comparison

Various polymorphs of triethylenetetramine salts may be formed by theinvention. Infrared analysis of a triethylenetetramine disuccinateproduced by an embodiment of the invention is set forth in FIG. 7.Infrared peaks were located at wavenumbers 3148, 1645, 1549, 1529, 1442,1370, 1311, 1271, 1221, 1172, 1152, 1085, 1052, 1033, 1003, 955, 922,866, and 827. Characteristic peaks of interest to identify thetriethylenetetramine disuccinate include 3148, 1549, 1529, 1442, 1370,and 1311 corresponding to —OH, —NH₂, —NH₂, —O, —CH₂, CH₂ respectively.DSC analysis of a triethylenetetramine disuccinate, with onset/peakmelting points of 180.05/179.91° C. is shown in FIG. 8.

At least one kinetic and one thermodynamic polymorph oftriethylenetetramine dihydrochloride may be formed by a process of theinvention. Infrared analysis of the thermodynamic polymorph (the Form Itriethylenetetramine dihydrochloride polymorph) is shown in FIG. 9.Infrared peaks are located at 825, 855, 925, 996, 1043, 1116, 1221,1300, 1328, 1367, 1401, 1457, 1476, 1503, 1557, 1619, 1640, 2705, 2833,2859, 2902, and 3216. DSC analysis of Form I triethylenetetraminedihydrochloride is shown in FIG. 10. Form I triethylenetetraminedihydrochloride has onset/peak melting points of 121.96/122.78° C.Infrared analysis of Form II triethylenetetramine dihydrochloride isshown in FIG. 11. Infrared peaks are located at wavenumbers 769, 807,866, 891, 965, 989, 1039, 1116, 1158, 1205, 1237, 1352, 1377, 1395,1438, 1456, 1519, 1610, 2102, 2419, 2468, 2533, 2611, 2667, 2743, 2954,2986, 3050, 3157, 3276, and 3298. DSC analysis of Form IItriethylenetetramine dihydrochloride has onset/peak melting points of116.16/116.76° C., as shown in FIG. 12.

Formation of Form I triethylenetetramine dihydrochloride versusformation of Form II triethylenetetramine dihydrochloride may bemediated by altered reaction conditions including cooling rate, presenceof seeding crystals, and number of equivalents of concentratedhydrochloric acid reacted with the free triethylenetetramine to formtriethylenetetramine dihydrochloride. In one embodiment, mixture of oneequivalent of triethylenetetramine with about 1.92 equivalents ofconcentrated hydrochloric acid in the presence of Form I seed crystalsyields Form I triethylenetetramine dihydrochloride.

Example 9

This Example demonstrates the reduction of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile (5) tointermediate 2-[3-(2-Amino-ethyl)-2-phenyl-imidazolidin-1-yl]-ethylamine(8). As shown in Scheme 9, intermediate (8) is treated with an acid toform a triethylenetetramine salt. The reduction is effected by LiAlH₄,the acid used is hydrochloric acid and the tetrahydrochloride salt oftriethylenetetramine is produced.

Example 10

This Example demonstrates preparation of triethylenetetramine vianitrile reduction using a Boc-protected intermediate. In the first stepof the reaction potassium carbonate (101.2 g, 0.71 mol, 2.0 eq) wassuspended in acetonitrile (200 mL). Ethylenediamine (21.34 g, 0.355 mol,1.0 eq) was added. The suspension was cooled to 5-10° C. and a solutionof chloroacetonitrile (57.02 g, 0.746 mol, 2.1 eq) in acetonitrile (40mL) was added. The mixture was stirred overnight at 25° C., with caretaken to prevent the reaction temperature from exceeding 35° C.Thin-layer chromatography showed almost complete conversion to thedinitrile intermediate [2-(cyanomethyl-amino)-ethylamino]-acetonitrile.The yellow suspension was cooled to 5° C. and Boc₂O (162.7 g, 0.746 mol,2.1 eq) was added in portions. After one hour, thin layer chromatographyshowed complete conversion to Boc-protected dinitrile,[2-(tert-Butoxycarbonyl-cyanomethyl-amino)-ethyl]-cyanomethyl-carbamicacid tert-butyl ester. The suspension was filtered and the filter cakewas washed with acetonitrile (800 mL). The filtrate was concentratedunder reduced pressure at 40° C. The crude brown solid,[2-(tert-Butoxycarbonyl-cyanomethyl-amino)-ethyl]-cyanomethyl-carbamicacid tert-butyl ester (122.8 g, 103%), was crystallized from from ethylacetate/methylcyclohexane (180 mL/600 mL). 62.83 g of crystallineBoc-protected dinitrile[2-(tert-Butoxycarbonyl-cyanomethyl-amino)-ethyl]-cyanomethyl-carbamicacid tert-butyl ester was obtained as a white solid. The mother liquorwas concentrated and again crystallized from ethylacetate/methylcyclohexane (100 mL/180 mL). 26.07 g of crystallineproduct was obtained as a white solid. A third crystallization yielded8.92 additional grams, resulting in a total yield of 81.3% ofBoc-protected dinitrile[2-(tert-Butoxycarbonyl-cyanomethyl-amino)-ethyl]-cyanomethyl-carbamicacid tert-butyl ester.

The Boc-protected dinitrile was then converted to Boc-protected diamine,(2-amino-ethyl)-{2-[(2-amino-ethyl)-tert-butoxycarbonyl-amino]-ethyl}-carbamicacid tert-butyl ester. The dinitrile (13.83 g, 0.0408 mol) was dissolvedin ethanol (150 mL) and NH₃ (25% in water) (12 mL). To the solution wasadded Raney nickel (12.8 g). The mixture was set under a hydrogenatmosphere (4-5 bar) for 15 hours. Thin-layer chromatography showedalmost complete conversion. The mixture was filtered and the solid waswashed with ethanol (350 mL). The filtrate was concentrated to drynessand the Boc-protected diamine(2-amino-ethyl)-{2-[(2-amino-ethyl)-tert-butoxycarbonyl-amino]-ethyl}-carbamicacid tert-butyl ester was obtained as 14.07 g of a white solid, a 99.5%yield.

The Boc-protected diamine(2-amino-ethyl)-{2-[(2-amino-ethyl)-tert-butoxycarbonyl-amino]-ethyl}-carbamicacid tert-butyl ester (2.83 g, 8.16 mmol) was dissolved in isopropanol(18 mL) and a mixture of concentrated HCl (5.5 mL, 48.9 mmol, 6 eq) inisopropanol (5.5 mL) was added. The mixture was heated to 70° C. for 30minutes. The resulting suspension was cooled to 20-25° C. and filtered.The solid was washed with TBME (12 mL) and dried on a rotary evaporator.2.16 g of triethylenetetramine tetrahydrochloride was obtained as awhite solid.

Example 11

Triethylenetetramine synthesis was accomplished through the use of a[2-(cyanomethyl-amino)-ethylamino]-acetonitrile intermediate (2)prepared by reaction of ethylenediamine (1). To prevent decomposition,the [2-(cyanomethyl-amino)-ethylamino]-acetonitrile intermediate (2) wasconverted to a Boc-derivative (i.e., protected by tert-butoxycarbonylgroups) as shown in Scheme 10.

Alkylation was performed by adding a solution of 2.2 equivalents ofchloroacetonitrile in acetonitrile to a mixture of 1 equivalent ofethylenediamine (1) and 2 equivalents of K₂CO₃ in acetonitrile overabout 30 minutes at 25° C. The reaction was complete within about 21hours. Upon complete conversion 1.2 equivalents of Boc₂O were added tothe reaction mixture. Thin-layer chromatography showed completeconversion to the Boc-protected intermediate (3), a[2-(tert-butoxycarbonyl-cyanomethyl-amino)-ethyl]-cyanomethyl-carbamicacid tert-butyl ester, after about 1 hour at 20° C. Work-up andcrystallization from ethyl acetate/methylcyclohexane ( 3/10) gaveBoc-protected intermediate (3) as a white solid in 81% yield.

Subsequent reaction of the Boc-protected dinitrile intermediate (3) wasperformed as shown in Scheme 11. A solution of the Boc-protecteddinitrile intermediate (3) in aqueous ethanol/NH₃ was hydrogenated inthe presence of Raney-nickel. A hydrogen atmosphere of 4-5 bar was usedand the reaction was allowed to run for about 15 hours at 20-25° C.Work-up and concentration to dryness afforded intermediate (4), a(2-amino-ethyl)-{2-[(2-amino-ethyl)-tert-butoxycarbonyl-amino]-ethyl}-carbamicacid tert-butyl ester, in 99% yield.

Heating a mixture of intermediate (4) and isopropanol with concentratedHCl to 70° C. for about 30 minutes cleaved the Boc-groups, as shown inScheme 2. The tetrahydrochloride salt of triethylenetetramine wasisolated by filtration in 91% yield. Presence of triethylenetetraminetetrahydrochloride salt was verified by NMR.

Example 12

This Example demonstrates the preparation of3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile. Equipment usedincluded a 160 L reactor, a 50 L nutsch, and a rotary vaporator equippedwith a 10 L round-bottom flask. The reactor was purged with nitrogengas. The scrubber was charged with about 30 L of a solution of 10% NaOH.The reactor was charged with about 120.3 mol of ethylenediaminedihydrochloride, then charged with about 238.2 mol of KCN. The reactorwas purged with nitrogen. The reaction was charged with about 60 L ofwater, and the internal temperature of the reactor dropped to about 9°C.

The reaction mixture was stirred for about 90 minutes at a maximumtemperature of about 30° C., until a temperature of about 20° C. wasreached, at which point the pH of the mixture was about 9.42. A 36%solution of formaldehyde in water (about 19.85 kg of the solution) wasadded with cooling over about 90 minutes. During the addition internalreaction temperature was maintained between about 20° C. and about 29°C. The pH of the reaction increased from about 9.12 to about 10.28. Thereaction mixture was stirred for about 2.5 hours at a range of about 20°C. to about 25° C.

Within thirty minutes of completion of addition of formaldehyde, a 43.6%solution of NaH₂PO₄ in water (about 13.1 L of solution) was added to thereaction at an internal reaction temperature of about 10° C. to about11° C. The pH of reaction mixture dropped from about 10.79 to about6.22. About 31 L of n-butyl alcohol were added to the reaction mixtureat an internal temperature of about 11° C. Within about thirty minutesof addition of n-butyl alcohol, about 120.3 moles of benzaldehyde wereadded to the reaction mixture at an internal temperature of about 11° C.to about 12° C. After about half of the benzaldehyde had been added,about 0.29 moles of3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile seed crystalswere added. During the addition the pH dropped from about 6.22 to about6.01. The reaction mixture was cooled to about −4° C. within about 5hours then stirred at about −4° C. for about 8.5 hours.

The nutsch was purged with hydrogen, and the reaction mixture wasfiltered. The filter cake was washed with about 47 L of deionized waterat about 15° C. in two portions, then washed with about 36 L of n-butylalcohol at about −3° C. in two portions, then washed with about 36 L ofisopropyl alcohol at about −2° C. in two portions. Crude(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile was transferredin four flasks and dried on a rotary evaporator at a temperature ofabout 40° C. and pressure less than or equal to about 20 mbar, producingabout 6.46 kg of (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrileat about 99.6% purity, about 8.12 kg(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile at about 99.2%purity, about 5.87 kg(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile at about 99.2%purity, and about 0.83 kg(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile at about 95.9%purity. All product was a white powder, and all purity measurements wereconducted by gas chromatography. Total yield of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile was about 21.28kg, which was about 78.5% of maximum amount possible given the amount ofstarting materials used.

Example 13

This Example demonstrates the preparation ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine.Equipment used was the same as that used in Example 10, above.

The reactor was purged with nitrogen. The reactor was charged with about5.00 kg of (3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile andpurged with nitrogen. About 22 L of THF was charged into the reactor,and the solution was transferred to a feeding vessel. The reactor waswashed with about 13 L of THF, and that solution was also transferred tothe feeding vessel.

The reactor was charged with about 46.2 kg of LiAlH₄ in 4% THE solution,which was cooled to an internal temperature of about 2.3° C. About 1.418kg of methanol and about 27 L of THF were mixed in another feedingvessel, and within about 20 minutes the mixture was added to the reactorat an internal temperature of about 0 to about 22° C. The solution wasthen heated to an internal temperature of about 40° C.

Within about three hours the solution of(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile in THF was addedat an internal temperature of about 40° C. A suspension was formed, andthe reaction mixture was stirred at about 40° C. for about one hour. Thereaction mixture was then cooled to an internal temperature of about 22°C. within about 30 minutes.

In process controls no longer detected(3-cyanomethyl-2-phenyl-imidazolidin-1-yl)-acetonitrile in solution. Thereaction mixture was further cooled to about 4° C., and within about onehour about 6.9 L of a solution of 4% NaOH was added. During addition thetemperature increased from about 4° C. to about 29° C. and hydrogen gasevolved. The reaction mixture was stirred at an internal temperature ofabout 10° C. overnight.

The nutsch was again purged with nitrogen, the suspension was filtered,and the filter cake was washed with THF. About 113 L of filtrateremained, and to this was added about 5.17 kg of benzaldehyde at 25° C.About 108 L THF was distilled from the solution at a temperature betweenabout 13 to about 30° C., with a pressure between about 100 and 200mbar.

About 40 L of isopropanol was added at about 40° C., and an additional 9L of solvent was distilled off at an internal temperature between about30 to about 37° C., under a pressure of about 90 to about 125 mbar. Thereaction mixture was cooled to an internal temperature of about 3° C.within about 30 minutes. About 7 g of seed crystals ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminewas added to the mixture at a temperature of about 30° C., and thereaction mixture was stirred at that temperature for about 30 minutes.The reaction mixture was then cooled to about −5° C. and stirredovernight.

The nutsch was again purged with nitrogen, the suspension was filtered,and the filter cake was washed with about 15 L of cold (about 0° C.)isopropanol. The product,benzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine,was dried in 2 round-bottom flasks on a rotary evaporator at atemperature of about 40° C. and a pressure less than or equal to 20mbar. Flask 1 gave about 2.864 kg ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of about 99.06 area % by gaschromatography. Flask 2 gave about 3.195 kg ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineas an off-white solid with a purity of about 99.24 area % by gaschromatography. Total yield ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminebased on starting materials was about 66.9%.

Example 14

This Example describes the preparation of triethylenetetraminedisuccinate. Reaction equipment was the same as that used in Examples 12and 13, above.

The reactor was purged with nitrogen. About 2.51 kg ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineand about 2.89 kg of succinic acid were added to a reactor. The reactorwas again purged with nitrogen. About 13 L of water were added to thereactor, and the mixture was heated to about 60° C. and stirred forabout 5 minutes. The mixture was cooled to about 20° C. About 13 L oftert-butylmethylether were added to the reaction mixture and stirred forabout 5 minutes, forming a biphasic mixture, with benzaldehyde in theorganic phase. The phases were separated and the organic layerdiscarded.

About 63 L of isopropanol were added to the aqueous phase at an internaltemperature of about 20° C. within about 20 minutes. After about 25 Lhad been added, about 8 g of triethylenetetramine disuccinate seedcrystals were added and addition of isopropanol continued. The reactionmixture was stirred at about 20° C. for about 70 minutes, and an about13 L of isopropanol were added to the mixture.

The reaction mixture was cooled to about 0° C. to facilitatecrystallization and stirred overnight. Triethylenetetramine disuccinateprecipitated from the isopropanol solution. The nutsch was purged withnitrogen, and the suspension was filtered. The filter cake was washedwith about 13 L of isopropanol. The product was then washed with about 6L of tert-butylmethylether, then dried on a rotary vaporator at atemperature of about 40° C. and a pressure less than or equal to about20 mbar.

The product, triethylenetetramine disuccinate, was an off-white solidproduced in an amount of about 2.098 kg, with a purity of about 100 area% by gas chromatography. Yield was about 89.9%.

Example 15

This Example describes an additional preparation of triethylenetetraminedisuccinate. Reaction equipment was the same as that used in Examples12, 13, and 14, above.

The reactor was purged with nitrogen. About 2.65 kg ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amineand about 3.05 kg of succinic acid were added to a reactor. The reactorwas again purged with nitrogen. About 13 L of water were added to thereactor, and the mixture was heated to about 59° C. and stirred forabout 5 minutes. The mixture was cooled to about 20° C. About 13 L oftert-butylmethylether were added to the reaction mixture and stirred forabout 13 minutes, forming a biphasic mixture, with benzaldehyde in theorganic phase. The phases were separated and the organic layerdiscarded.

About 66 L of isopropanol were added to the aqueous phase at an internaltemperature of about 20° C. within about 21 minutes. After about 25 Lhad been added, about 9 g of triethylenetetramine disuccinate seedcrystals were added and addition of isopropanol continued. The reactionmixture was stirred at about 19 to 22° C. for about 65 minutes, and anabout 13 L of isopropanol was added to the mixture.

The reaction mixture was cooled to about 5° C. to facilitatecrystallization and stirred overnight. Triethylenetetramine disuccinateprecipitated from the isopropanol solution. The nutsch was purged withnitrogen, and the suspension was filtered. The filter cake was washedwith about 14 L of isopropanol. The product was washed with about 7 L oftert-butylmethylether, then dried on a rotary vaporator at a temperatureof about 40° C. and a pressure less than or equal to about 20 mbar.

The product, triethylenetetramine disuccinate, was an off-white solidproduced in an amount of about 2.266 kg, with a purity of about 100 area% by gas chromatography. Yield was about 91.7%.

Example 16

This Example describes a further additional preparation oftriethylenetetramine disuccinate. Reaction equipment was the same asthat used in Examples 10, 11, 12, and 13 above.

1 equal mole ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-aminewas added to the reactor, which was purged before and after withnitrogen. Isopropanol was added to the reactor, followed by the additionof of water. The reaction mixture was then heated to 40° C. and thentransferred into a second reactor via an inline-filter. 4 equal moles ofsuccinic acid was added to the first reactor which was then purged withnitrogen, followed by addition of methanol and stirring for 40 minutesor until a clear solution was obtained. The succinic acid solution wasthen transferred to the second reactor via the inline-filter, at thesame time of transferal the same volume of methanol added was distilledoff at ET=50° C., p=>100 mbar and internal temperature (IT)=22-28° C.The mixture in the second reactor is stirred for about one hour at 40°C., cooled to IT=0° C. and then stirred overnight at this sametemperature. The resultant suspension was filtered and isopropanol addedthrough the inline filter, followed by the addition of tert-butyl methylether (TBME) via the inline-filter. The resultant triethylenetetraminedisuccinate was then dried in a PROVATECH-dryer (ET=40° C., p≦20 mbar)in two portions.

The triethylenetetramine disuccinate was then re-crystallized to removebenzaldehyde. Dried disuccinate salt was placed back into the reactor,purged with nitrogen and water added and heated to IT=50° C. Methanolwas inline-filtered and added to the reaction mixture followed by water.The mixture was then heated to IT=55° C. at which point a clear solutionwas obtained. Methanol was inline-filtered and added to the reactionmixture over 25 minutes at IT=55° C. and then cooled to 0° C. for 30minutes. The resultant suspension was filtered and methanol wasinline-filtered to the reactor. The resultant triethylenetetraminedisuccinate was dried in the PROVATECH-dryer (ET=40° C., p<20 mbar).This resulted in a white to off white solid, purity of 98.84 area % (IonChromatography) with a yield of 93.1%.

Example 17

This Example describes the preparation of triethylenetetraminetetrahydrochloride by a process of the invention. Reaction equipment wassimilar to that used in Example 15.

The reactor was purged with nitrogen and charged with about 9.46 kg ofbenzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1}-ethyl)-amine.The reactor was again purged with nitrogen, and about 15 L of water wereintroduced to the reactor. About 14.5 L of a 32% solution of HCl wereadded to the aqueous suspension with cooling between temperatures ofabout 17 to about 20° C. within 26 minutes.

The reaction mixture was cooled at about 17° C. for about fifteenminutes, until a clear solution was obtained. About 30 L oftert-butylmethyl ether were added to the reactor at a temperature ofabout 20° C., and the mixture was stirred at about that temperature forabout 10 minutes. A biphasic mixture was formed. The organic layer wasdiscarded.

Within 1.5 hours about 89 L of isopropanol were added to the aqueoussolution at about 18-22° C. The suspension was stirred at about 23° C.for about 15 minutes. The nutsch was purged with nitrogen and thesuspension was filtered. The filter cake was washed with about 30 L ofisopropanol, washed with about 30 L of tert-butylmethyl ether, and againwashed with about 30 L of tert-butylmethyl ether. The cake was dried ona rotary evaporator at about 40° C., at a pressure less than or equal toabout 20 mbar in three lots.

Triethylenetetramine tetrahydrochloride was produced. In Flask 1, about2.19 kg of an off-white solid with a purity of about 98.5 area % by gaschromatography and about 100.0% by chloride titration assay wasobtained. In Flask 2, about 1.94 kg of an off-white solid with a purityof about 98.2 area % by gas chromatography and about 99.9% by chloridetitration assay was obtained. In Flask 3, about 2.08 kg of an off-whitesolid with a purity of about 98.3 area % by gas chromatography and about102.9% by chloride titration assay was obtained. This corresponded to atotal yield of about 92.3%.

Example 18

This Example describes the preparation of triethylenetetraminedihydrochloride by a process of the invention. Equipment used was thesame as that in Example 16.

The reactor was purged with nitrogen and charged with about 6.00 kg oftriethylenetetramine tetrahydrochloride. The reactor was purged againwith nitrogen, then about 60 L of ethanol were added. About 14.56 kg ofNaOMe (about 30.5% in methanol) were diluted with about 25 L of ethanol.The diluted solution was added to the suspension in the reactor vesselat a temperature of about 19 to about 22° C. within 10 minutes. Thefeeding vessel of the NaOMe solution was washed with about 4.8 L ofethanol into the reactor.

The suspension was stirred at about 20° C. for about 11 minutes. About24 L of tert-butylmethyl ether were charged into the reactor, and thesuspension was stirred at about 20° C. for about three hours. Thesuspension was filtered. The filter cake was washed with about 24 L of amixture of tert-butylmethyl ether and ethanol. The filtrate wasconcentrated in the reactor vessel by distillation of about 130 L ofsolvent at about 19 to about 22° C., with a pressure of about 47 toabout 120 mbar.

The reactor was charged with about 30 L of tert-butylmethyl ether. Themixture was stirred at about 23° C. for about 15 minutes. The suspensionwas filtered, and the filter cake was washed with about 6 liters oftert-butylmethyl ether. The filtrate was filtered into the reactorvessel through an inline filter, and the inline filter was rinsed withabout 5 L of tert-butylmethyl ether. The filtrate was concentrated inthe reactor vessel by distillation of about 32 L of solvent at aninternal temperature of about 17 to 24° C. and a pressure of about 45 to170 mbar.

The solution was cooled to about 20° C. within about 7 minutes. About 20L of ethanol were added through an inline filter, and the solution wascooled to an internal temperature of about 0° C. within about 16minutes. About 4.46 kg of HCl was added, with cooling, through an inlinefilter at an internal temperature of between about 0 and 15° C. withinabout 30 minutes.

The suspension was stirred at an internal temperature of about 15 toabout 22° C. for about 10 minutes, then heated to about 48° C. for about1 hour until a clear solution was obtained. About 133 L of ethanol wereadded through an inline filter. About 10 g of seed crystals of athermodynamic polymorph of triethylenetetramine dihydrochloride wereadded at an internal temperature of about 32° C. The reaction mixturewas stirred for about thirty minutes at about 29 to about 32° C. until asuspension formed. The suspension was cooled to an internal temperatureof about 3° C. within 5 hours, then stirred at about 3° C. for about 11hours. The nutsch was purged with nitrogen, and the suspension wasfiltered.

The filter cake was washed with about 15 L of ethanol, then washed withabout 15 L of tert-butylmethyl ether. The filter was dried on a rotaryvaporator at a temperature of about 40° C. and a pressure less than orequal to about 20 mbar. About 3.62 kg of triethylenetetraminedihydrochloride was obtained as a yellowish solid with a purity of about100 area % by TLC. This corresponded to a yield of about 80.4%.

Example 19

This Example illustrates, triethylenetetramine tetramaleate,triethylenetetramine tetrafumarate and triethylenetetramine disuccinate,the synthesis of these salts according to the synthetic schemesdescribed above. Crystals of X-ray quality for triethylenetetraminetetramaleate were grown by slow evaporation of a supersaturated solutionof triethylenetetramine tetramaleate in water. The triethylenetetraminedisuccinate and triethylenetetramine tetrafumarate were grown by slowevaporation of a solution of 12.58 mg triethylenetetramine disuccinateand 7.42 mg triethylenetetramine tetrafumarate in a water/ethanolmixture (1:1, 2 ml) over a period of 3 weeks. Crystal structure data(Tables 3-5) was obtained by single crystal x-ray diffractionmeasurement. For comparison, x-ray powder diffraction measurements weredone with the accordant powder material.

FIG. 13 shows a crystal structure of triethylenetetramine disuccinateanhydrate. Based on the resolved structure, the composition ratio oftriethylenetetramine:succinate was confirmed to be 1:2. Thetriethylenetetramine and the succinate molecules formed alternatinglayers which interact via strong hydrogen bonds. The positively chargedtriethylenetetramines do not interact with each other. FIG. 14 shows thetriethylenetetramine molecule surrounded by eight succinate molecules.Six of these succinate molecules formed one very strong hydrogen bondbetween one of the negatively charged O-atoms and the H-atom of theprotonated NH- or NH₂-groups. The other two succinate molecules formedtwo hydrogen bonds between their negatively charged O-atom and an H-atomof a protonated NH-group and another H-atom of the protonated NH₂-group.Water molecules are not necessary to complete the coordination sphere ofthe triethylenetetramine molecule. Each succinate molecule iscoordinated via six hydrogen bonds to four triethylenetetraminemolecules (FIG. 14 II). FIGS. 15 and 16 represent two X-ray powderdiffraction patterns obtained from independently synthesizedtriethylenetetramine disuccinate powder material. Crystal quality may beenhanced by an additional recrystallisation.

TABLE 3A Crystallographic and refinement data for triethylenetetraminedisuccinate formula sum C₁₄ H₃₄ N₄ O₈ formula weight 386.44 measurementtemperature 84(2) K measurement device Bruker SMART CCD wavelength0.71076 Å (Mo-Kα-radiation) crystal system monoclinic space group C 2/c(no. 15) unit cell dimensions a = 14.059(5) Å b = 9.169(5) Å c =13.647(5) Å β = 92.47(0)° cell volume 1757.56(130) Å³ Z 4 density,calculated 1.007 g/cm³ R_(All) 0.043 absorption coefficient μ 0.077 mm⁻¹F(000) 584 θ range 2.65-25.66° h_(min), h_(max); k_(min), k_(max);l_(min), l_(max) −17, 16; −6, 11; −14, 16 reflections measured 4899[R(int) = 0.0322] independent reflections 1653 observed reflections [I >2s(i)] 1455 data/restraints/parameters 1653/0/118 Goodness-of-fit at F²1.053 R indices [I > 2 sigma(I)] R1 = 0.0366, wR2 = 0.0948 R indices(all data) R1 = 0.0431, wR2 = 0.0987 largest diff. peak/hole0.266/−0.260 e · A⁻³

TABLE 3B Triethylenetetramine disuccinate atomic coordinates andisotropic displacement parameters (in Å²) x y z U_(eq) C1 0.29558(10)0.26210(16) 0.31932(10) 0.0126(3) H1A 0.25420 0.34680 0.31880 0.01500H1B 0.27720 0.19900 0.37230 0.01500 C2 0.2803(1) 0.18094(16) 0.22253(10)0.0122(3) H2A 0.33450 0.11790 0.21290 0.01500 H2B 0.22430 0.119800.22590 0.01500 C3 0.2663(1) 0.19924(16) 0.04178(9) 0.0124(3) H3A0.22310 0.11710 0.04510 0.01500 H3B 0.32940 0.16200 0.03010 0.01500 C40.0209(1) 0.46728(16) 0.12805(9) 0.0113(3) C5 −0.0467(1) 0.33720(16)0.11701(10) 0.0129(3) H5A −0.08810 0.35300 0.05930 0.01500 H5B −0.086500.33540 0.17330 0.01500 C6 −0.00004(10) 0.18713(16) 0.10792(10)0.0134(3) H6A 0.04010 0.18650 0.05180 0.01600 H6B 0.03980 0.167400.16620 0.01600 C7 −0.07602(10) 0.06870(16) 0.09545(10) 0.0114(3) N10.39566(8) 0.31058(14) 0.33899(8) 0.0117(3) H1C 0.40020 0.35710 0.396200.01800 H1D 0.41260 0.37040 0.29140 0.01800 H1E 0.43390 0.23320 0.341100.01800 N2 0.26795(8) 0.28179(14) 0.13633(8) 0.0109(3) H2C 0.213100.33160 0.14060 0.01300 H2D 0.31610 0.34660 0.13740 0.01300 O1−0.01814(7) 0.59176(11) 0.12409(7) 0.0154(3) O2 0.10919(7) 0.44737(11)0.14249(7) 0.0153(3) O3 −0.12598(8) 0.06534(12) 0.01693(7) 0.0188(3) O4−0.08767(7) −0.01952(12) 0.16553(7) 0.0178(3)

TABLE 3C Triethylenetetramine disuccinate anisotropic displacementparameters (in Å²) U₁₁ U₂₂ U₃₃ U₁₂ U₁₃ U₂₃ C1 0.0120(7) 0.0130(8)0.0129(7) 0.0010(6) 0.0011(5) 0.0010(6) C2 0.0135(7) 0.0099(7) 0.0130(7)−0.0003(5) −0.0007(5) 0.0017(5) C3 0.0133(7) 0.0113(7) 0.0125(7)0.0000(5) −0.0007(5) −0.0024(6) C4 0.0132(7) 0.0121(8) 0.0087(6)0.0002(6) 0.0016(5) −0.0012(5) C5 0.0106(7) 0.0123(8) 0.0158(7)−0.0010(6) 0.0007(5) −0.0002(5) C6 0.0125(7) 0.0122(8) 0.0154(7)−0.0013(6) 0.0011(5) 0.0006(5) C7 0.0128(7) 0.0092(7) 0.0123(6)0.0019(5) 0.0018(5) −0.0011(5) N1 0.0136(6) 0.0111(6) 0.0104(5)0.0010(5) −0.0008(4) −0.0006(4) N2 0.0108(6) 0.0103(6) 0.0115(6)−0.0003(5) −0.0005(4) 0.0000(5) O1 0.0139(5) 0.0106(6) 0.0216(5)0.0007(4) −0.0008(4) −0.0006(4) O2 0.0107(5) 0.0131(6) 0.0219(5)0.0001(4) 0.0001(4) −0.0026(4) O3 0.0241(6) 0.0185(6) 0.0132(5)−0.0079(4) 0.0044(4) 0.0033(4) O4 0.0206(6) 0.0174(6) 0.0150(5)−0.0066(4) −0.0032(4) 0.0055(4)

Example 20

FIG. 17 shows a crystal structure with the composition oftriethylenetetramine tetramaleate.2H₂O obtained fromtriethylenetetramine tetramaleate. X-ray diffraction measurements of thecrystals confirmed the 1:4 ratio of triethylenetetramine:maleate presentin the crystal based on the analysis of the powder material. FIG. 18shows that the obtained crystal structure is characterized by a layerstructure which contained alternating layers of triethylenetetramine andmaleate molecules. FIG. 18 shows that the triethylenetetramine moleculeis surrounded by eight maleate and two water molecules. Each maleatemolecule possesses a mono negative charge and forms a strong hydrogenbond between the negatively charged O-atom of the maleate and the H-atomof the protonated NH- or NH₂-groups. The H-atom of the COOH-group of themaleate molecule also forms hydrogen bonds to anothertriethylenetetramine molecule, with slightly longer bond distances. EachH-atom of the protonated NH- or NH₂-groups forms a hydrogen bond to theO-atom of one maleate molecule, except for one H-atom of each NH₃⁺-group at each end of the molecule, which form a hydrogen bond to anadditional water molecule. Thus, these two water molecules complete thecoordination sphere of the NH₃ ⁺-groups of the triethylenetetraminemolecule. As seen in FIG. 18, the triethylenetetramine molecules areonly connected to each other via the additional water molecules (seeFIG. 18, II) within the layers. Triethylenetetramine and maleate areconnected via the hydrogen bonds described above. Inside the layer ofthe maleate there is no significant interaction between the molecules.FIG. 19 represents an x-ray powder diffraction pattern obtained from asynthesized triethylenetetramine. Crystal quality may be enhanced by anadditional recrystallisation.

TABLE 4A Crystallographic and refinement data for triethylenetetraminetetramaleate dihydrate formula sum C₂₂ H₃₈ N₄ O₁₈ formula weight 645.56measurement temperature 83 K measurement device Bruker SMART CCDwavelength 0.71069 Å (Mo-Kα-radiation) crystal system monoclinic spacegroup P 2/c (no. 13) unit cell dimensions a = 13.261(5) Å b = 9.342(5) Åc = 11.266(5) Å β = 91.01(0)° cell volume 1395.46(110) Å³ Z 4 density,calculated 1.229 g/cm³ R_(All) 0.047 absorption coefficient μ 0.130 mm⁻¹F(000) 664 θ range 1.54-26.41° h_(min), h_(max); k_(min), k_(max);l_(min), l_(max) −16, 16; −8, 11; −14, 14 reflections measured 8095[R(int) = 0.0198] independent reflections 2860 observed reflections [I >2s(i)] 2457 data/restraints/parameters 2860/0/224 Goodness-of-fit at F²1.103 R indices [I > 2 sigma(I)] R1 = 0.0937, wR2 = 0.0375 R indices(all data) R1 = 0.0998, wR2 = 0.0475 largest diff. peak/hole0.279/−0.217 e · A⁻³

TABLE 4B Triethylenetetramine tetramaleate atomic coordinates andisotropic displacement parameters (in Å²) x y z U_(eq) C1 −0.1177(1)0.3688(2) 0.0564(1) 0.0176(3) C2 −0.1132(1) 0.4687(2) 0.1600(1)0.0209(3) H2 −0.1111 0.4243 0.2339 0.02500 H2A 0.1153 −0.1147 0.05470.02900 H2B 0.1366 0.0378 0.0565 0.02900 H2C 0.0887 −0.0293 0.15730.02900 C3 −0.1116(1) 0.6113(2) 0.1633(1) 0.0208(3) H3 −0.1067 0.65020.2391 0.02500 C4 −0.1166(1) 0.7188(2) 0.0656(1) 0.0203(3) C5 0.6193(1)0.6463(2) 0.5558(1) 0.0178(3) C6 0.6208(1) 0.5439(2) 0.4540(1) 0.0187(3)H6 0.6160 0.5861 0.3794 0.02200 C7 0.6279(1) 0.4015(2) 0.4531(1)0.0191(3) H7 0.6272 0.3604 0.3779 0.02300 C8 0.6368(1) 0.2971(2)0.5531(1) 0.0180(3) C9 0.2656(1) 0.0548(2) 0.2328(1) 0.0173(3) H9A0.2704 0.1405 0.1846 0.02100 H9B 0.2145 0.0712 0.2920 0.02100 C100.2343(1) −0.0703(2) 0.1549(1) 0.0178(3) H10A 0.2834 −0.0843 0.09320.02100 H10B 0.2316 −0.1570 0.2022 0.02100 C11 0.4516(1) 0.0112(2)0.2127(1) 0.0180(3) H11 0.4417(12) −0.0783(18) 0.1690(14) 0.016(4) H120.4482(12) 0.0925(18) 0.1592(15) 0.018(4) H13 −0.0348(16) 0.158(2)0.2007(18) 0.049(6) H14 −0.1340(19) 0.550(3) −0.047(2) 0.071(8) H150.6350(18) 0.477(3) 0.668(2) 0.066(8) H16 0.3586(12) −0.0514(19)0.3391(15) 0.019(4) H17 0.3759(13) 0.108(2) 0.3430(16) 0.026(4) N10.3645(1) 0.0276(1) 0.2934(1) 0.0157(3) N2 0.1336(1) −0.0412(1)0.1004(1) 0.0191(3) O1 −0.1086(1) 0.2382(1) 0.0779(1) 0.0220(2) O2−0.1305(1) 0.4170(1) −0.0485(1) 0.0240(3) O3 −0.1077(1) 0.8469(1)0.0914(1) 0.0271(3) O4 −0.1314(1) 0.6768(1) −0.0434(1) 0.0244(3) O50.6440(1) 0.1687(1) 0.5282(1) 0.0229(3) O6 0.6372(1) 0.3416(1) 0.6613(1)0.0284(3) O7 0.6315(1) 0.6009(1) 0.6624(1) 0.0274(3) O8 0.6062(1)0.7750(1) 0.5322(1) 0.0216(2) O9 0.0000 0.0994(2) 0.2500 0.0194(3)

TABLE 4C Triethylenetetramine tetramaleate anisotropic displacementparameters (in Å²) U₁₁ U₂₂ U₃₃ U₁₂ U₁₃ U₂₃ C1 0.0167(7) 0.0198(7)0.0163(7) −0.0006(5) 0.0003(5) −0.0013(6) C2 0.0273(8) 0.0216(8)0.0137(7) 0.0024(6) 0.0007(6) 0.0011(6) C3 0.0256(7) 0.0218(8) 0.0148(7)0.0007(6) −0.0002(5) −0.0028(6) C4 0.0196(7) 0.0205(8) 0.0208(7)−0.0001(6) −0.0005(5) 0.0005(6) C5 0.0185(7) 0.0190(7) 0.0160(7)−0.0017(5) 0.0015(5) −0.0011(6) C6 0.0250(7) 0.0193(7) 0.0118(7)−0.0002(6) 0.0015(5) 0.0019(5) C7 0.0246(7) 0.0209(8) 0.0118(6)0.0005(6) 0.0014(5) −0.0013(6) C8 0.0185(7) 0.0186(7) 0.0168(7)−0.0009(5) 0.0001(5) 0.0005(5) C9 0.0185(7) 0.0170(7) 0.0163(7)0.0009(5) 0.0004(5) −0.0003(5) C10 0.0195(7) 0.0178(7) 0.0162(7)0.0010(5) −0.0001(5) −0.0007(6) C11 0.0198(7) 0.0218(8) 0.0124(6)0.0002(6) 0.0020(5) −0.0010(6) N1 0.0194(6) 0.0141(6) 0.0137(6)−0.0006(4) 0.0010(5) 0.0000(5) N2 0.0218(6) 0.0191(6) 0.0164(6)0.0003(5) −0.0015(5) −0.0022(5) O1 0.0285(6) 0.0170(5) 0.0204(5)0.0014(4) −0.0032(4) −0.0013(4) O2 0.0364(6) 0.0211(6) 0.0142(5).0007(5) −0.0028(4) −0.0007(4) O3 0.0353(6) 0.0182(6) 0.0278(6)−0.0019(5) −0.0026(5) −0.0006(5) O4 0.0355(6) 0.0201(6) 0.0176(5)−0.0018(4) −0.0022(4) 0.0025(4) O5 0.0322(6) 0.0173(5) 0.0193(5)−0.0001(4) 0.0007(4) 0.0011(4) O6 0.0511(7) 0.0216(6) 0.0125(5)0.0005(5) −0.0017(5) 0.0006(4) O7 0.0479(7) 0.0203(6) 0.0137(5)0.0006(5) −0.0035(5) −0.0016(4) O8 0.0283(6) 0.0168(5) 0.0198(5)−0.0004(4) 0.0020(4) −0.0018(4) O9 0.0219(7) 0.0177(7) 0.0186(7) 0.00000−0.0018(6) 0.00000

Example 21

The crystal structure of the triethylenetetramine tetrafumarate isrelated to the structure of triethylenetetramine tetramaleate. Thecrystal structure data (FIG. 20) revealed a compound composition oftriethylenetetramine tetrafumarate.4H₂O. Analysis of the structureconfirmed the 1:4 ratio of triethylenetetramine:fumarate as four watermolecules were found. The triethylenetetramine molecule interacts viastrong hydrogen bonds between the H-atoms of the protonated NH- andNH₂-groups and the O-atoms of eight fumarate molecules, except for oneH-atom of each NH₃ ⁺-group at each end of the molecule which forms anadditional bond to a water molecule. In this case, it appears that twowater molecules may be necessary to complete the coordination sphere ofthe NH₃ ⁺-groups of the triethylenetetramine molecule. See FIG. 21. Eachfumarate molecule is connected to four different triethylenetetraminemolecules via two short and two long hydrogen bonds. FIG. 20 shows thattriethylenetetramine tetrafumarate forms a layered structure oftriethylenetetramine and fumarate molecules. In contrast totriethylenetetramine tetramaleate, the triethylenetetramine layers oftriethylenetetramine tetrafumarate are shifted along the c axis of c/2against each other. Within the layers, the triethylenetetraminemolecules are only connected to each other via the additional watermolecules. The layers of triethylenetetramine and fumarate are alsoconnected via hydrogen bonds. Within the fumarate layers, the moleculesare arranged in pairs with alternating orientations and there areadditional water molecules located between these molecules. Thereappears to be no significant interactions between the fumarate and thewater molecules. In the case of triethylenetetramine tetrafumarate, acomparison of the powder pattern of triethylenetetramine tetrafumaratepowder material with calculated data of the triethylenetetraminetetrafumarate.4H₂O crystal structure, which contains four additionalwater molecules, shows that there is a complete conformity of allreflexes between these two compounds. FIG. 22 represents an x-ray powderdiffraction pattern obtained from a synthesized triethylenetetraminetetrafumarate. Crystal quality may be enhanced by an additionalrecrystallisation. A comparison with the analytical data shows that thecompound is hygroscopic and the NMR, IR and DSC results also indicatethat the compound contains water.

TABLE 5A Crystallographic and refinement data for triethylenetetraminetetrafumarate tetrahydrate formula sum C₄₄ H₂₀ N₈ O₄₄ formula weight1364.68 measurement temperature 84(2) K measurement device Bruker SMARTCCD wavelength 0.71073 Å (Mo-Kα-radiation) crystal system orthorhombicspace group P m n a (no. 53) unit cell dimensions a = 13.9031(3) Å b =7.9589(2) Å c = 14.6554(3) Å cell volume 1621.67(6) Å³ Z 1 density,calculated 1.397 g/cm³ R_(All) 0.075 absorption coefficient μ 0.130 mm⁻¹F(000) 692 θ range 2.02-26.39° h_(min), h_(max); k_(min), k_(max);l_(min), l_(max) −17, 17; −8, 9; −18, 12 reflections measured 9146[R(int) = 0.0573] independent reflections 1728 observed reflections [I >2s(i)] 1541 data/restraints/parameters 1728/0/139 Goodness-of-fit at F²1.187 R indices [I > 2 sigma(I)] R1 = 0.0678, wR2 = 0.1457 R indices(all data) R1 = 0.0754, wR2 = 0.1502 largest diff. peak/hole0.455/−0.381 e · A⁻³

TABLE 5B Triethylenetetramine tetrafumarate atomic coordinates andisotropic displacement parameters (in Å²) x y z U_(eq) N1 0 0.5928(3)0.1267(2) 0.0252(6) H1A 0 0.67640 0.08640 0.0380 H1B −0.05230 0.529900.11880 0.0380 H1C 0.05230 0.52990 0.11880 0.0380 N2 0 0.5907(3)0.3820(2) 0.0232(6) H2A −0.05230 0.65600 0.38910 0.0280 H2B 0.052300.65600 0.38910 0.0280 C1 0 0.6630(4) 0.2203(2) 0.0256(7) H1D −0.056500.73250 0.22920 0.0310 H1E 0.05650 0.73250 0.22920 0.0310 C2 0 0.5204(4)0.2885(2) 0.0242(7) H2C −0.05650 0.45090 0.27970 0.0290 H2D 0.056500.45090 0.27970 0.0290 C3 0 0.4584(4) 0.4541(3) 0.0245(7) H3A 0.0559(19)0.393(3) 0.4448(18) 0.033(7) C4 0.22620(16) 0.3189(3) 0.12220(19)0.0284(6) C5 0.27015(17) 0.1484(3) 0.11941(18) 0.0290(6) H5A 0.336700.14060 0.11490 0.0350 C6 0.22071(16) 0.0093(3) 0.12297(18) 0.0255(5)H6A 0.15410 0.01590 0.12780 0.0310 C7 0.26664(16) 0.8402(3) 0.11968(18)0.0264(5) O1 0.20734(12) 0.7174(2) 0.12499(15) 0.0370(5) H1F 0.245(3)0.585(6) 0.120(3) 0.119(18) O2 0.35426(12) 0.8256(2) 0.11207(17)0.0407(6) O3 0.28558(12) 0.4403(2) 0.12399(16) 0.0413(6) O4 0.13799(13)0.3371(2) 0.12186(17) 0.0464(6) O5 0 0.1729(4) 0.0124(2) 0.0322(6) H5B 00.095(10) 0.004(7) 0.14(4) H5C 0.053(4) 0.209(6) 0.052(3) 0.022(13) O6 01.0804(8) 0.2523(9) 0.134(6) O7 0 1.0541(11) 0.4360(14) 0.143(10)

TABLE 5C Triethylenetetramine tetrafumarate anisotropic displacementparameters (in Å²) U₁₁ U₂₂ U₃₃ U₁₂ U₁₃ U₂₃ N1 0.0184(13) 0.0167(13)0.0406(17) 0.00000 0.00000 −0.0023(12) N2 0.0156(12) 0.0107(12)0.0432(17) 0.00000 0.00000 0.0006(12) C1 0.0218(15) 0.0148(14)0.0402(19) 0.00000 0.00000 −0.0030(14) C2 0.0177(14) 0.0127(14) 0.042(2)0.00000 0.00000 −0.0007(14) C3 0.0199(15) 0.0108(14) 0.043(2) 0.000000.00000 0.0002(14) C4 0.0195(11) 0.0147(10) 0.0509(16) 0.0006(9)0.0005(10) −0.0024(10) C5 0.0193(11) 0.0146(11) 0.0530(16) 0.0024(8)0.0042(11) −0.0002(11) C6 0.0193(10) 0.0150(11) 0.0423(14) 0.0013(9)−0.0041(10) 0.0013(10) C7 0.0206(11) 0.0139(10) 0.0448(14) 0.0005(8)−0.0016(10) 0.001(1) O1 0.0189(8) 0.0110(8) 0.0812(15) −0.0003(6)0.0009(9) 0.0014(8) O2 0.0189(9) 0.0139(8) 0.0894(16) 0.0012(6)0.0012(9) 0.0056(9) O3 0.0204(9) 0.0110(8) 0.0925(17) 0.0002(6)0.0010(9) 0.0019(9) O4 0.0207(9) 0.0143(8) 0.1042(18) 0.0015(7)0.0032(10) −0.0078(10) O5 0.0287(13) 0.0282(14) 0.0395(16) 0.000000.00000 0.0051(12) O6 0.065(5) 0.038(4) 0.299(18) 0.00000 0.00000−0.009(6) O7 0.036(5) 0.030(5) 0.36(3) 0.00000 0.00000 −0.014(8)

Patents, patent applications, publications, scientific articles, books,web sites, and other documents and materials referenced or mentionedherein are indicative of the levels of skill of those skilled in the artto which the inventions pertain. Each such referenced document andmaterial is hereby incorporated by reference to the same extent as if ithad been incorporated by reference in its entirety individually or setforth or reprinted herein in its entirety. Additionally, all claims inthis application, and all priority applications, including but notlimited to original claims, are hereby incorporated in their entiretyinto, and form a part of, the written description of the invention.Applicants reserve the right to physically incorporate into thisspecification any and all materials and information from any suchpatents, applications, publications, scientific articles, web sites,electronically available information, and other referenced materials ordocuments. Applicants reserve the right to physically incorporate intoany part of this document, including any part of the writtendescription, and the claims referred to above including but not limitedto any original claims.

The inventions have been described broadly and generically herein. Eachof the narrower species and subgeneric groupings falling within thegeneric disclosure also form part of these inventions. This includes thegeneric description of each invention which hereby include, includingany claims thereto, a proviso or negative limitation removing oroptionally allowing the removal of any subject matter from the genus,regardless of whether or not the excised materials or options werespecifically recited or identified in haec verba herein, and all suchvariations form a part of the original written description of theinventions. In addition, where features or aspects of an invention aredescribed in terms of a Markush group, the invention shall be understoodthereby to be described in terms of each and every, and any, individualmember or subgroup of members of the Markush group.

Although the invention has been described in terms of synthesis oftriethylenetetramines and triethylenetetramine salts, it should berecognized that the routes, steps, and intermediates described in thedisclosure are applicable to the synthesis of polyethylenepolyamines andpolyethylenepolyamine salts of the formulaNH₂CH₂(CH₂NHCH₂CH₂NHCH₂)_(n)CH₂NH₂ through synthesis, protection, andreduction as described herein of dinitrile intermediates of the generalformula NC(CH₂NHCH₂CH₂NHCH₂)_(n)CN, where n is greater than or equal to1.

The inventions illustratively described and claimed herein can suitablybe practiced in the absence of any element or elements, limitation orlimitations, not specifically disclosed herein or described herein asessential. Thus, for example, the terms “comprising,” “including,”“containing,” “for example”, etc., shall be read expansively and withoutlimitation. The term “including” means “including but not limited to.”The phrase “for example” is not limited to or by the items that followthe phrase. All references to things “known in the art” include allthose things and equivalents and substitutes, whether now known or laterdiscovered.

In claiming their inventions, the inventors reserve the right tosubstitute any transitional phrase with any other transitional phrase,and the inventions shall be understood to include such substitutedtransitions and form part of the original written description of theinventions. Thus, for example, the term “comprising” may be replacedwith either of the transitional phrases “consisting essentially of” or“consisting of.”

The methods and processes illustratively described herein may besuitably practiced in differing orders of steps. They are notnecessarily restricted to the orders of steps indicated herein or in theclaims.

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural reference unless the context clearly dictatesotherwise.

Under no circumstances may the patent be interpreted to be limited tothe specific examples or embodiments or methods specifically disclosedherein. Under no circumstances may the patent be interpreted to belimited by any statement made by any Examiner or any other official oremployee of the Patent and Trademark Office unless such statement wasspecifically and without qualification or reservation expressly adoptedby Applicants in a responsive writing specifically relating to theapplication that led to this patent prior to its issuance.

The terms and expressions employed herein have been used as terms ofdescription and not of limitation, and there is no intention in the useof such terms and expressions, or any portions thereof, to exclude anyequivalents now know or later developed, whether or not such equivalentsare set forth or shown or described herein or whether or not suchequivalents are viewed as predictable, but it is recognized that variousmodifications are within the scope of the invention claimed, whether ornot those claims issued with or without alteration or amendment for anyreason. Thus, it shall be understood that, although the presentinvention has been specifically disclosed by preferred embodiments andoptional features, modifications and variations of the inventionsembodied therein or herein disclosed can be resorted to by those skilledin the art, and such modifications and variations are considered to bewithin the scope of the inventions disclosed and claimed herein.

Specific methods and compositions described herein are representative ofpreferred embodiments and are exemplary and not intended as limitationson the scope of the invention. Other objects, aspects, and embodimentswill occur to those skilled in the art upon consideration of thisspecification, and are encompassed within the spirit of the invention asdefined by the scope of the claims. Where examples are given, thedescription shall be construed to include but not to be limited to onlythose examples. It will be readily apparent to one skilled in the artthat varying substitutions and modifications may be made to theinvention disclosed herein without departing from the scope and spiritof the invention, and from the description of the inventions, includingthose illustratively set forth herein, it is manifest that variousmodifications and equivalents can be used to implement the concepts ofthe present invention without departing from its scope. A person ofordinary skill in the art will recognize that changes can be made inform and detail without departing from the spirit and the scope of theinvention. The described embodiments are to be considered in allrespects as illustrative and not restrictive. Thus, for example,additional embodiments are within the scope of the invention and withinthe following claims.

1. A compound of formula (II),

wherein each R of formula (II) is independently methyl carbamate, ethylcarbamate, benzyl carbamate, tert-butyl carbamate, tert-butyloxycarbonyl(Boc), cyclohexanone, 2,2,6,6-tetramethyl cyclohexanone, anthrone, analkyl group, an aryl group, or an aromatic alkyl group, or a compound offormula (VI),

wherein each R of formula (VI) is independently methyl carbamate, ethylcarbamate, benzyl carbamate, tert-butyl carbamate, tert-butyloxycarbonyl(Boc), cyclohexanone, 2,2,6,6-tetramethyl cyclohexanone, anthrone, analkyl group, an aryl group, or an aromatic alkyl group.
 2. The compoundof claim 1, wherein R is tert-butyloxycarbonyl (Boc).
 3. The compound ofclaim 1, wherein the compound is crystallized.
 4. The compound of claim1, wherein the compound is substantially pure.
 5. The compound of claim3, wherein the compound is substantially pure.